树鼩肝癌发生过程中的差异蛋白质组学研究及Prx家族的表达验证
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摘要
肝细胞性肝癌(hepatocellular carcinoma, HCC)是诊治困难、预后极差的一种恶性肿瘤,全球三大肿瘤死因之一,1990年全世界共有54.1万新发的肝癌病例,50.1万人死于肝癌,其中31.8万(58.5%)新发病例和29.3万(58.8%)肝癌死亡发生在中国。HCC的病因目前认为主要与乙型肝炎病毒(HBV)感染及食品中黄曲霉毒素B_1(AFB_1)污染有关。和其它多数肿瘤一样,HCC的形成是一个多步骤、多阶段、多因素参与的过程,传统的单基因研究模式限制了HCC相关因素的研究发展。近年来迅速发展的蛋白质组学研究,能动态、整体、定量地考察疾病发生发展过程中全部蛋白质种类和数量的变化,对于探讨疾病机理、寻找疾病诊断的特异性标志物和药物治疗的靶标来说,是一种有效的高通量的研究模式,可获得一些传统手段无法得到的新蛋白标志物、新关键分子,极大地丰富了诊断标志物的选择组合及复杂致病机理的阐明。运用该研究策略,建立并优化了适合本实验的双向凝胶电泳(2-DE)技术平台,并利用树鼩可以在实验诱癌过程中经受多次剖腹肝活检手术而长期存活的特点,本研究对HCC形成过程中不同时期肝组织的蛋白质组改变进行动态观察。对AFB_1和AFB_1+人HBV诱发树鼩肝癌发生不同阶段的差异表达蛋白进行筛选,找出在HCC形成过程中一些有潜在作用的差异蛋白质,并对其中的差异蛋白Prx家族在肝癌组织和癌旁组织进行验证,为探讨人HCC发生机理提供重要科学依据。
第一部分人工繁育树鼩的生理指标与正常人的对比
本部分的研究目的是通过对人工繁育的成年树鼩进行实验室指标的检测,以之与正常人参考值进行比较,分析二者间的关系,探讨树鼩用于研究人疾病动物模型的可行性及优越性。
选择人工繁育饲养的成年树鼩22只,同一天内抽取树鼩的抗凝全血和分离血清,随即进行相关实验室指标的检测。结果为,血常规检测,红细胞总数8.19×10~(12)/L,高于正常人参考值,白细胞总数2.2×10~9/L,低于正常人参考值;每份全血标本涂片和染色后镜检观察,树鼩各种血细胞及血小板形态都与正常人相似;白细胞分类计数,树鼩各种细胞百分比与正常人相近;血清标本生化十项指标检测,仅树鼩肌酐一项结果为16.63μmol/L,低于正常人参考值下限44μmol/L,树鼩的血清谷丙转氨酶77.92U/L、谷草转氨酶179.9 U/L分别高于正常人参考值上限40 U/L和45 U/L,其余七项结果符合正常人参考值范围,包括尿素、球蛋白、血清总蛋白、血清白蛋白、血清γ-谷氨酰基转移酶、C-反应蛋白及类风湿因子;检测HBsAg为0.022 ng /ml±0.005,均为阴性。
以上各实验室指标检测结果提示,树鼩作为接近人的低等灵长类动物,适合应用于研究人类疾病的动物模型,为进一步利用树鼩模型研究HCC发生机理提供了实验依据。
第二部分树鼩肝癌蛋白质组学双向凝胶电泳技术的建立及优化
本部分研究的目的是通过建立并优化树鼩肝癌蛋白质组学的双向凝胶电泳技术,提高其分辨率和重复性,并利用该平台进行AFB_1和AFB_1+人HBV诱发树鼩肝癌形成过程中差异表达蛋白质组学的研究。
实验以AFB_1和AFB_1+HBV诱发树鼩肝癌形成过程中各时期的肝组织标本,通过对传统的双向凝胶电泳技术的程序及有关环节进行反复改进,尤其对其中的关键因素环节,如样品的处理(单纯的液氮下研磨、液氮下研磨再剧烈振荡)、蛋白定量方法(Lowery法和改良Bradford法)、上样方式(杯上样、泡胀上样)、等电聚焦程序和凝胶染色方法(考马斯亮蓝R-250染色、传统银染、质谱兼容银染)等,进行了一系列的优化。
实验结果通过优化获得了较为理想的电泳图谱:图谱蛋白点数明显增多(1228个vs 856个);图谱分辨率提高,水平条带及纵向拖尾现象明显减少。同时,2-DE的重复性也有大幅度提高:对同组样品在三次不同实验中所得蛋白质斑点数目分别为1241、1192和1216个,IEF方向平均偏差1.73±0.99 mm,SDS-PAGE方向平均偏差1.11±0.75mm。本技术平台的建立为后续工作奠定了基础。
第三部分树鼩肝癌发生过程中差异表达蛋白质组学的研究
本部分的研究目的是应用2-DE和MALDI-TOF-MS等蛋白质组学技术,比较分析AFB_1与AFB_1+人HBV诱发树鼩肝癌发生过程中肝组织蛋白质表达谱的差异,寻找在树鼩肝癌发生过程中起重要作用的关键蛋白质分子,为探讨人HCC发生机理提供重要科学依据。
本研究利用树鼩可以在实验诱癌过程中经受多次剖腹肝活检手术而长期存活的特点,对HCC形成过程中不同时期肝组织、癌前各时期的肝活检组织、肝癌和癌旁组织,以及各同期对照组织蛋白质组改变进行动态观察。由于细胞或组织蛋白质组的复杂性,应用2-DE分离总蛋白时,为保证结果的准确性及精确度,本研究共12个实验组:①AFB_1实验组(A组),A组45周肝活检组织(A45w)、A组75周肝活检组织(A75w)、A组肝癌组织(ACa)、A组癌旁组织(ACp);②AFB_1+HBV实验组(C组),C组45周肝活检组织(C45w)、C组肝癌组织(CCa)、C组癌旁组织(CCp);③HBV实验组(D组),D组45周肝活检组织(D45w)、D组105周肝活检组织(D105w);④正常对照组(E组),E组45周肝活检组织(E45w)、E组75周肝活检组织(E75w)、E组105周肝活检组织(E105w),每组6例组织样本的混合总蛋白分别进行3次2-DE以保证试验的重复性。蛋白质均用13cm pH 3-10非线性IPG胶条,采用泡胀上样方式进行等电聚焦电泳,共获得36块2D胶图谱。
各组蛋白表达谱中蛋白多集中分布于pI 4-8间的区域,用ImageMaster 2D Platinum 5.0软件将凝胶分类别进行点的匹配分析。每组3块胶,各组平均检测到的蛋白点数如下: A45w组1255±43个,A75w组1197±46个,ACa组1198±50个,ACp组1205±89个;C45w组1233±30个,CCa组1257±35个,CCp组1244±39个;D45w组1100±79个,D105w组1115±61个;E45w组1249±51个,E75w组1179±62个,E105w组1208±36个。筛选出差异2倍及其以上并且在半数胶以上中均存在的蛋白点共206个。
从相应的考马斯亮蓝染色的制备胶中挖取差异倍数较大且蛋白斑点较清晰的差异蛋白点160个,进行MALDI-TOF-MS/MS分析。为保证取点的准确性,每个点均从三块胶中挖取后分别进行酶解鉴定,取质谱鉴定结果一致者。鉴定出的差异蛋白点共142个,95种蛋白,包括peroxiredoxin家族(PrxⅠ、PrxⅡ、PrxⅢ、PrxⅣ、PrxⅥ)、细胞色素C氧化酶、谷胱甘肽过氧化物酶Ⅰ、硫氧还蛋白、细胞色素B5、酮己糖激酶、不均一核糖核蛋白、磷酸丙糖异构酶、尼克酰胺N-甲基转移酶、转移抑制基因NM23-H1、翻译控制肿瘤蛋白等。
第四部分差异蛋白Prx家族在树鼩及人HCC组织中的表达验证
由于第三部分筛选鉴定的结果显示, peroxiredoxin家族(PrxⅠ、PrxⅡ、PrxⅢ、PrxⅣ)在AFB_1诱发树鼩HCC形成过程中,多个成员蛋白均出现明显差异表达:PrxⅠ、PrxⅡ、PrxⅢ在癌组织中表达均较癌前组织明显上调,PrxⅣ则在癌组织中较癌旁组织明显上调,并且由于Prx的生物学功能广泛并与多种肿瘤相关,因此选择该组蛋白作为进一步研究目标。
本部分的研究目的是对差异蛋白Prx家族在树鼩HCC发生不同阶段的蛋白水平和mRNA水平的表达差异,以及在人肝癌组织和癌旁组织间的表达差异进行再验证,进一步解析该蛋白的生物学功能,探讨Prx家族蛋白在肝癌发生发展中的作用以及其作用机制。
应用Western Blot技术,本部分结果证实PrxⅠ、PrxⅡ、PrxⅢ蛋白表达水平在AFB_1诱发树鼩HCC组织(ACa)明显高于癌前组织(A45w),略高于正常对照组织(E45w),PrxⅣ蛋白表达水平亦发生上调,但差异未达2倍。人HCC组织验证结果,PrxⅠ、Ⅱ、Ⅲ、Ⅳ的表达量都略高于癌旁组织,但升高倍数均未达到2倍。运用RT-PCR技术检测,PrxⅠ、Ⅱ、Ⅲ、Ⅳ在树鼩和人的mRNA表达水平升高趋势与蛋白表达水平相同。Prx家族蛋白有望成为一类有应用价值的肝癌标志物蛋白,有可能应用于HCC的诊断、治疗和预后评价。
结论
1、树鼩作为低等灵长类动物,各项实验室指标检测结果接近正常人,为其用于建立研究人类疾病的动物模型进一步提供了支持依据。
2、通过对2-DE相关技术的摸索,建立和优化了适合于本实验的树鼩肝癌蛋白质组学双向凝胶电泳技术平台,为后续研究奠定了基础。
3、与对照组各时期的正常肝组织相比,树鼩HCC发生过程中各时期肝组织表现出有一定差异性的蛋白表达谱,表明HCC的发生发展是一个综合的多因素参与的过程。鉴定出的142个,95种差异表达蛋白质,可能参与肝癌的发生发展。
4、Prx家族蛋白有望成为一类有应用价值的肝癌标志物蛋白,有可能应用于HCC的诊断、治疗和预后评价。
潜在应用价值
1、树鼩作为低等灵长类动物,适合应用于研究人类疾病的动物模型。
2、建立了树鼩肝癌蛋白质组学双向凝胶电泳技术平台。
3、筛检出的差异蛋白有可能成为肝癌发生的诊断标志物及预后判断指标,丰富了肝癌诊疗指标的选择与组合。
4、Prx可以作为一种有应用价值的测定肝癌发生发展的标志物。
创新点
1.建立并优化了树鼩肝癌蛋白质组学双向凝胶电泳技术平台。
2.利用树鼩可以在实验诱癌过程中经受多次剖腹肝活检手术而长期存活的特点,对HCC形成过程中不同时期肝组织的蛋白质组改变进行动态分析,筛选出一批与肝癌发生不同阶段相关的差异蛋白。
3.分析了与特定病因因素:AFB_1、AFB_1 +人HBV诱发HCC相关的差异表达蛋白。
4.分析了肝组织感染HBV不同时期的差异表达蛋白。
5.首次证实差异蛋白PrxⅠ、PrxⅡ、PrxⅢ、PrxⅣ在AFB_1诱发的树鼩肝癌发生过程中出现上调。
Hepatocellular carcinoma (HCC), of which the morbidity and mortality are in the forefront among the overall tumors worldwide, is one of the malignancies with difficult in early diagnosis and extremely poor prognosis. Hepatitis B virus (HBV) infection and aflatoxin B_1 (AFB_1) exposure are recognized as the main factors for HCC. The formation of HCC is a multistep, multistage and multifactor process like most other tumors. The conventional strategy of research on single gene has been challenged by the rapidly progress of proteomics, which can be utilized to observe the dynamical changes of nature and quantity of total protein during progression of disease. Proteomics can be used to find new protein markers and key molecules and therefore is an effective and high-throughout method for exploring mechanism of disease and for searching targets of diagnosis or therapy. By optimizing the technical platform of two-dimensional electrophoresis (2-DE), in the present study we observed dynamically the differentially expressed proteins of liver tissue that were taken at different time points during the hepatocarcinogenesis of tree shrew (Tupaia belangeri chinensis), a small animal species belonged to low Primates evolutionarily. By taking the advantages that the animal can survive repeated liver biopsies during the experiment and can be infected with HBV, the present study aimed at screening differential expressed proteins in various stages of hepatocarcinogenesis induced by AFB_1 or AFB_1+HBV. Subsequently, one group of the screened proteins, peroxiredoxin family (Prxs), was further confirmed by Western blot and RT-PCR on the tissues samples form tree shrews as well as from HCC patients. The entire study included four parts as following.
Part 1. Normal physiological laboratory value of tree shrew
To further confirm the feasibility and superiority of utilizing tree shrew as animal model for studying human disease, this part is to detect the physiological laboratory values of adult tree shrew bred artificially, and to comparer the values with human normal reference.
Whole blood sample and separated serum were collected from twenty two adult tree shrews bred artificially in this lab, and then all the laboratory values were determined. Blood routine examination showed that RBC count was 8.19×10~(12)/L, which was higher than human normal reference value. Tree shrew WBC count was 2.2×10~9/L, which was lower than human normal reference. Microscopic examination of whole blood smear from tree shrew showed the morphology of blood cells and platelet were similar to that of human. WBC differential count showed that each cell percentage was near to that of human. Assay of ten biochemical values of serum samples showed that seven values including urea, globulin, serum total protein, serum albumin, serumγ-GT, C-reactive protein and rheumatoid factor accorded with the range of human reference. While serum creatinine of tree shrew as 16.63μmol/L was lower than 44μmol/L of human reference, serums GPT as 77.92 U/L and GOT as 179.9 U/L were higher than the upper limit of human reference of 40 U/L and 45 U/L respectively. The result of HBsAg examination showed all of the animals were negative, with the value of 0.022 ng/ml±0.005.
Analysis of these laboratory values mentioned above suggests that tree shrew, as a low grade primate related closely to humankind, is suitable as animal model for studying human disease.
Part 2. Establish and optimize 2-DE technique for proteomic study on tree shrew liver samples
For achieving a high quality on resolution and a steady result, effort was made to establish and optimize the 2-DE technique that should be adaptive for proteomic analysis on tree shrew liver samples. The conventional protocol of 2-DE was modified, especially on some critical steps such as sample processing (grinding and homogenating), quantitative methods of protein (Lowery method and modified Bradford method), loading modes (cup-loading and swelling-loading), IEF procedure, gel stain methods (Coomassie brilliant blue R-250 stain, traditional silver stain, MS compatible silver stain), and etc.
The protein spots acquired from 2-DE gel by the modified method increased greatly (1228 vs 856), along with the enhanced spot-resolution, as well as with the decreased horizontal band and vertical tail. The protein spots from same sample that run repeatedly three times of 2-DE were 1241, 1192 and 1216 respectively, with the mean shiftings as 2.0±0.56mm at IEF direction and 2.3±0.61mm at SDS-PAGE direction. Establishment of this technical platform provided the foundation for following research.
Part 3. Proteomic study on differential expression during tree shrew’s hepatocarcinogenesis
To search the critical protein molecules responsible for tree shrew hepatocarcinogenesis, and to explore the mechanism of HCC. This part compared and analyzed protein profiles of tree shrew HCC induced by AFB_1 and AFB_1+HBV, by 2-DE and MALD1-TDF MS proteomic techniques.
Twelve categories of liver samples were collected from differently treated animals at different time points during the experiment. Samples from the animals in group A that were treated with AFB_1 and developed HCC at late stage of the experiment, named A45w, A75w, ACa and ACp respectively, referring the samples were biopsies at 45th week (A45w) or 75th week (A75w) of the experiment before HCC appeared, and the HCC samples (ACa) or HCC-surrounding samples (ACp) were from the identical animals. Similarly, the samples from the animals in group C that were not only treated with AFB_1 but also infected with HBV and developed HCC at late stage of the experiment were named C45w, CCa and CCp respectively. The samples from the animals in group D that were only infected with HBV, had no HCC developed until the experiment ended at 165th week, were named D45w and D105w respectively. The samples from the control animals in group E that treated nothing else but only breed normally were named E45w, E75w and E105w respectively. The mixed total protein from samples in each category was assayed repeatedly with 2-DE method. Thirteen centimeters non-linear IPG gel strip with pH 3-10 was adopted. The IEF electrophoresis was carried out by applying the swelling sample, and pictures from 36 blocks of 2-DE gels were acquired.
Profile of proteins was similar among the different categories of sample as most protein spots distributed inside the field of pI 4-8. ImageMaster 2-DE Platimum 5.0 software was utilized to carry out analysis for each well-matched gel dot between the comparable categories. The mean detected numbers of protein spot in each categories were as follows: 1255±43 in A45w, 1197±46 in A75w, 1198±50 in ACa, 1205±89 in ACp; 1233±30 in C45w, 1257±35 in CCa, 1244±39 in CCp; 1100±79 in D45w, 1115±61 in D105w; 1249±51 in E45w, 1179±62 in E75w, 1208±36 in E105w. Two hundred and six protein points which had double or more density and appeared in more than fifty percent gels were screened out.
One hundred sixty protein spots with different expression were dug out from the corresponding prepared Coomassie brilliant blue stain gels and one hundred forty-two spots were identified by MALDI-TOF MS. To assure the accuracy of blots sampling, each spot dug out from three blocks of gel was identified and only identical data in MS was selected. The identified differential proteins included the members of Peroxiredoxin family (PrxⅠ, PrxⅡ, PrxⅢ, and PrxⅣ), cytochrome C oxidase, glutathione peroxidaseⅠ, thioredoxin, cytochrome B5, ketohexose kinase, heterogeneous ribose nucleoprotein, phosphotriose isomerase, nicotinamide N-methyl transferase, metastasis suppressed gene NM23-H1, translation controlled tumor protein, and others.
Part 4. Verify tests on differential expression of peroxiredoxins among liver samples from tree shrews and from HCC patients
The members in peroxiredoxin family (PrxⅠ, PrxⅡ, PrxⅢand PrxⅣ), which were among the differentially expressed protein screened from Part 3 and were known had widespread biological functions concerning many other tumors, were studied further to confirm the 2-DE results that showed the expressions of PrxⅠ, PrxⅡ, and PrxⅢin HCC tissue were up-regulated greatly compared to precancerous tissue, and the expression of PrxⅣwas up-regulated prominently in HCC tissue compared to HCC-surrounding liver tissue. By Western blot and RT-PCR assays, the expression levels of PrxⅠ, PrxⅡ, PrxⅢand PrxⅣwere examined on the samples from tree shrew as well as from HCC patients.
Results confirmed that the expressions of PrxⅠ, PrxⅡ, PrxⅢ, and PrxⅣwere not only up-regulated in tree shrew HCC but also in human HCC tissue in both protein level and mRNA level, which indicated that the proteins in Prx family might play important role in hepatocarcinogenesis and could be hopefully applied as a biomarker for HCC diagnosis and/or treatment.
Conclusion
Tree shrew, a lower grade primate related closely with humankind evolutionarily, is proved suitable as animal model to research human disease because most of its laboratory values were found similar to that of human beings.
The modified 2-DE technical platform for proteomic analysis on liver tissues of tree shrews provides the foundation for research. Profiles of differentially expressed protein screened from the tissues taken at different time points during tree shrew hepatocarcinogenesis induced by AFB_1 or AFB_1+HBV suggests the progression of HCC is concerned with many factors. At least some of the proteins among the 142 identified ones might play important roles in the HCC initiation, development and/or progression. Particularly, proteins in Prx family might be applied as a biomarker for HCC diagnosis, treatment and/or prognosis-evaluation.
第一部分人工繁育树鼩的生理指标与正常人的对比
本部分的研究目的是通过对人工繁育的成年树鼩进行实验室指标的检测,以之与正常人参考值进行比较,分析二者间的关系,探讨树鼩用于研究人疾病动物模型的可行性及优越性。
选择人工繁育饲养的成年树鼩22只,同一天内抽取树鼩的抗凝全血和分离血清,随即进行相关实验室指标的检测。结果为,血常规检测,红细胞总数8.19×10~(12)/L,高于正常人参考值,白细胞总数2.2×10~9/L,低于正常人参考值;每份全血标本涂片和染色后镜检观察,树鼩各种血细胞及血小板形态都与正常人相似;白细胞分类计数,树鼩各种细胞百分比与正常人相近;血清标本生化十项指标检测,仅树鼩肌酐一项结果为16.63μmol/L,低于正常人参考值下限44μmol/L,树鼩的血清谷丙转氨酶77.92U/L、谷草转氨酶179.9 U/L分别高于正常人参考值上限40 U/L和45 U/L,其余七项结果符合正常人参考值范围,包括尿素、球蛋白、血清总蛋白、血清白蛋白、血清γ-谷氨酰基转移酶、C-反应蛋白及类风湿因子;检测HBsAg为0.022 ng /ml±0.005,均为阴性。
以上各实验室指标检测结果提示,树鼩作为接近人的低等灵长类动物,适合应用于研究人类疾病的动物模型,为进一步利用树鼩模型研究HCC发生机理提供了实验依据。
第二部分树鼩肝癌蛋白质组学双向凝胶电泳技术的建立及优化
本部分研究的目的是通过建立并优化树鼩肝癌蛋白质组学的双向凝胶电泳技术,提高其分辨率和重复性,并利用该平台进行AFB_1和AFB_1+人HBV诱发树鼩肝癌形成过程中差异表达蛋白质组学的研究。
实验以AFB_1和AFB_1+HBV诱发树鼩肝癌形成过程中各时期的肝组织标本,通过对传统的双向凝胶电泳技术的程序及有关环节进行反复改进,尤其对其中的关键因素环节,如样品的处理(单纯的液氮下研磨、液氮下研磨再剧烈振荡)、蛋白定量方法(Lowery法和改良Bradford法)、上样方式(杯上样、泡胀上样)、等电聚焦程序和凝胶染色方法(考马斯亮蓝R-250染色、传统银染、质谱兼容银染)等,进行了一系列的优化。
实验结果通过优化获得了较为理想的电泳图谱:图谱蛋白点数明显增多(1228个vs 856个);图谱分辨率提高,水平条带及纵向拖尾现象明显减少。同时,2-DE的重复性也有大幅度提高:对同组样品在三次不同实验中所得蛋白质斑点数目分别为1241、1192和1216个,IEF方向平均偏差1.73±0.99 mm,SDS-PAGE方向平均偏差1.11±0.75mm。本技术平台的建立为后续工作奠定了基础。
第三部分树鼩肝癌发生过程中差异表达蛋白质组学的研究
本部分的研究目的是应用2-DE和MALDI-TOF-MS等蛋白质组学技术,比较分析AFB_1与AFB_1+人HBV诱发树鼩肝癌发生过程中肝组织蛋白质表达谱的差异,寻找在树鼩肝癌发生过程中起重要作用的关键蛋白质分子,为探讨人HCC发生机理提供重要科学依据。
本研究利用树鼩可以在实验诱癌过程中经受多次剖腹肝活检手术而长期存活的特点,对HCC形成过程中不同时期肝组织、癌前各时期的肝活检组织、肝癌和癌旁组织,以及各同期对照组织蛋白质组改变进行动态观察。由于细胞或组织蛋白质组的复杂性,应用2-DE分离总蛋白时,为保证结果的准确性及精确度,本研究共12个实验组:①AFB_1实验组(A组),A组45周肝活检组织(A45w)、A组75周肝活检组织(A75w)、A组肝癌组织(ACa)、A组癌旁组织(ACp);②AFB_1+HBV实验组(C组),C组45周肝活检组织(C45w)、C组肝癌组织(CCa)、C组癌旁组织(CCp);③HBV实验组(D组),D组45周肝活检组织(D45w)、D组105周肝活检组织(D105w);④正常对照组(E组),E组45周肝活检组织(E45w)、E组75周肝活检组织(E75w)、E组105周肝活检组织(E105w),每组6例组织样本的混合总蛋白分别进行3次2-DE以保证试验的重复性。蛋白质均用13cm pH 3-10非线性IPG胶条,采用泡胀上样方式进行等电聚焦电泳,共获得36块2D胶图谱。
各组蛋白表达谱中蛋白多集中分布于pI 4-8间的区域,用ImageMaster 2D Platinum 5.0软件将凝胶分类别进行点的匹配分析。每组3块胶,各组平均检测到的蛋白点数如下: A45w组1255±43个,A75w组1197±46个,ACa组1198±50个,ACp组1205±89个;C45w组1233±30个,CCa组1257±35个,CCp组1244±39个;D45w组1100±79个,D105w组1115±61个;E45w组1249±51个,E75w组1179±62个,E105w组1208±36个。筛选出差异2倍及其以上并且在半数胶以上中均存在的蛋白点共206个。
从相应的考马斯亮蓝染色的制备胶中挖取差异倍数较大且蛋白斑点较清晰的差异蛋白点160个,进行MALDI-TOF-MS/MS分析。为保证取点的准确性,每个点均从三块胶中挖取后分别进行酶解鉴定,取质谱鉴定结果一致者。鉴定出的差异蛋白点共142个,95种蛋白,包括peroxiredoxin家族(PrxⅠ、PrxⅡ、PrxⅢ、PrxⅣ、PrxⅥ)、细胞色素C氧化酶、谷胱甘肽过氧化物酶Ⅰ、硫氧还蛋白、细胞色素B5、酮己糖激酶、不均一核糖核蛋白、磷酸丙糖异构酶、尼克酰胺N-甲基转移酶、转移抑制基因NM23-H1、翻译控制肿瘤蛋白等。
第四部分差异蛋白Prx家族在树鼩及人HCC组织中的表达验证
由于第三部分筛选鉴定的结果显示, peroxiredoxin家族(PrxⅠ、PrxⅡ、PrxⅢ、PrxⅣ)在AFB_1诱发树鼩HCC形成过程中,多个成员蛋白均出现明显差异表达:PrxⅠ、PrxⅡ、PrxⅢ在癌组织中表达均较癌前组织明显上调,PrxⅣ则在癌组织中较癌旁组织明显上调,并且由于Prx的生物学功能广泛并与多种肿瘤相关,因此选择该组蛋白作为进一步研究目标。
本部分的研究目的是对差异蛋白Prx家族在树鼩HCC发生不同阶段的蛋白水平和mRNA水平的表达差异,以及在人肝癌组织和癌旁组织间的表达差异进行再验证,进一步解析该蛋白的生物学功能,探讨Prx家族蛋白在肝癌发生发展中的作用以及其作用机制。
应用Western Blot技术,本部分结果证实PrxⅠ、PrxⅡ、PrxⅢ蛋白表达水平在AFB_1诱发树鼩HCC组织(ACa)明显高于癌前组织(A45w),略高于正常对照组织(E45w),PrxⅣ蛋白表达水平亦发生上调,但差异未达2倍。人HCC组织验证结果,PrxⅠ、Ⅱ、Ⅲ、Ⅳ的表达量都略高于癌旁组织,但升高倍数均未达到2倍。运用RT-PCR技术检测,PrxⅠ、Ⅱ、Ⅲ、Ⅳ在树鼩和人的mRNA表达水平升高趋势与蛋白表达水平相同。Prx家族蛋白有望成为一类有应用价值的肝癌标志物蛋白,有可能应用于HCC的诊断、治疗和预后评价。
结论
1、树鼩作为低等灵长类动物,各项实验室指标检测结果接近正常人,为其用于建立研究人类疾病的动物模型进一步提供了支持依据。
2、通过对2-DE相关技术的摸索,建立和优化了适合于本实验的树鼩肝癌蛋白质组学双向凝胶电泳技术平台,为后续研究奠定了基础。
3、与对照组各时期的正常肝组织相比,树鼩HCC发生过程中各时期肝组织表现出有一定差异性的蛋白表达谱,表明HCC的发生发展是一个综合的多因素参与的过程。鉴定出的142个,95种差异表达蛋白质,可能参与肝癌的发生发展。
4、Prx家族蛋白有望成为一类有应用价值的肝癌标志物蛋白,有可能应用于HCC的诊断、治疗和预后评价。
潜在应用价值
1、树鼩作为低等灵长类动物,适合应用于研究人类疾病的动物模型。
2、建立了树鼩肝癌蛋白质组学双向凝胶电泳技术平台。
3、筛检出的差异蛋白有可能成为肝癌发生的诊断标志物及预后判断指标,丰富了肝癌诊疗指标的选择与组合。
4、Prx可以作为一种有应用价值的测定肝癌发生发展的标志物。
创新点
1.建立并优化了树鼩肝癌蛋白质组学双向凝胶电泳技术平台。
2.利用树鼩可以在实验诱癌过程中经受多次剖腹肝活检手术而长期存活的特点,对HCC形成过程中不同时期肝组织的蛋白质组改变进行动态分析,筛选出一批与肝癌发生不同阶段相关的差异蛋白。
3.分析了与特定病因因素:AFB_1、AFB_1 +人HBV诱发HCC相关的差异表达蛋白。
4.分析了肝组织感染HBV不同时期的差异表达蛋白。
5.首次证实差异蛋白PrxⅠ、PrxⅡ、PrxⅢ、PrxⅣ在AFB_1诱发的树鼩肝癌发生过程中出现上调。
Hepatocellular carcinoma (HCC), of which the morbidity and mortality are in the forefront among the overall tumors worldwide, is one of the malignancies with difficult in early diagnosis and extremely poor prognosis. Hepatitis B virus (HBV) infection and aflatoxin B_1 (AFB_1) exposure are recognized as the main factors for HCC. The formation of HCC is a multistep, multistage and multifactor process like most other tumors. The conventional strategy of research on single gene has been challenged by the rapidly progress of proteomics, which can be utilized to observe the dynamical changes of nature and quantity of total protein during progression of disease. Proteomics can be used to find new protein markers and key molecules and therefore is an effective and high-throughout method for exploring mechanism of disease and for searching targets of diagnosis or therapy. By optimizing the technical platform of two-dimensional electrophoresis (2-DE), in the present study we observed dynamically the differentially expressed proteins of liver tissue that were taken at different time points during the hepatocarcinogenesis of tree shrew (Tupaia belangeri chinensis), a small animal species belonged to low Primates evolutionarily. By taking the advantages that the animal can survive repeated liver biopsies during the experiment and can be infected with HBV, the present study aimed at screening differential expressed proteins in various stages of hepatocarcinogenesis induced by AFB_1 or AFB_1+HBV. Subsequently, one group of the screened proteins, peroxiredoxin family (Prxs), was further confirmed by Western blot and RT-PCR on the tissues samples form tree shrews as well as from HCC patients. The entire study included four parts as following.
Part 1. Normal physiological laboratory value of tree shrew
To further confirm the feasibility and superiority of utilizing tree shrew as animal model for studying human disease, this part is to detect the physiological laboratory values of adult tree shrew bred artificially, and to comparer the values with human normal reference.
Whole blood sample and separated serum were collected from twenty two adult tree shrews bred artificially in this lab, and then all the laboratory values were determined. Blood routine examination showed that RBC count was 8.19×10~(12)/L, which was higher than human normal reference value. Tree shrew WBC count was 2.2×10~9/L, which was lower than human normal reference. Microscopic examination of whole blood smear from tree shrew showed the morphology of blood cells and platelet were similar to that of human. WBC differential count showed that each cell percentage was near to that of human. Assay of ten biochemical values of serum samples showed that seven values including urea, globulin, serum total protein, serum albumin, serumγ-GT, C-reactive protein and rheumatoid factor accorded with the range of human reference. While serum creatinine of tree shrew as 16.63μmol/L was lower than 44μmol/L of human reference, serums GPT as 77.92 U/L and GOT as 179.9 U/L were higher than the upper limit of human reference of 40 U/L and 45 U/L respectively. The result of HBsAg examination showed all of the animals were negative, with the value of 0.022 ng/ml±0.005.
Analysis of these laboratory values mentioned above suggests that tree shrew, as a low grade primate related closely to humankind, is suitable as animal model for studying human disease.
Part 2. Establish and optimize 2-DE technique for proteomic study on tree shrew liver samples
For achieving a high quality on resolution and a steady result, effort was made to establish and optimize the 2-DE technique that should be adaptive for proteomic analysis on tree shrew liver samples. The conventional protocol of 2-DE was modified, especially on some critical steps such as sample processing (grinding and homogenating), quantitative methods of protein (Lowery method and modified Bradford method), loading modes (cup-loading and swelling-loading), IEF procedure, gel stain methods (Coomassie brilliant blue R-250 stain, traditional silver stain, MS compatible silver stain), and etc.
The protein spots acquired from 2-DE gel by the modified method increased greatly (1228 vs 856), along with the enhanced spot-resolution, as well as with the decreased horizontal band and vertical tail. The protein spots from same sample that run repeatedly three times of 2-DE were 1241, 1192 and 1216 respectively, with the mean shiftings as 2.0±0.56mm at IEF direction and 2.3±0.61mm at SDS-PAGE direction. Establishment of this technical platform provided the foundation for following research.
Part 3. Proteomic study on differential expression during tree shrew’s hepatocarcinogenesis
To search the critical protein molecules responsible for tree shrew hepatocarcinogenesis, and to explore the mechanism of HCC. This part compared and analyzed protein profiles of tree shrew HCC induced by AFB_1 and AFB_1+HBV, by 2-DE and MALD1-TDF MS proteomic techniques.
Twelve categories of liver samples were collected from differently treated animals at different time points during the experiment. Samples from the animals in group A that were treated with AFB_1 and developed HCC at late stage of the experiment, named A45w, A75w, ACa and ACp respectively, referring the samples were biopsies at 45th week (A45w) or 75th week (A75w) of the experiment before HCC appeared, and the HCC samples (ACa) or HCC-surrounding samples (ACp) were from the identical animals. Similarly, the samples from the animals in group C that were not only treated with AFB_1 but also infected with HBV and developed HCC at late stage of the experiment were named C45w, CCa and CCp respectively. The samples from the animals in group D that were only infected with HBV, had no HCC developed until the experiment ended at 165th week, were named D45w and D105w respectively. The samples from the control animals in group E that treated nothing else but only breed normally were named E45w, E75w and E105w respectively. The mixed total protein from samples in each category was assayed repeatedly with 2-DE method. Thirteen centimeters non-linear IPG gel strip with pH 3-10 was adopted. The IEF electrophoresis was carried out by applying the swelling sample, and pictures from 36 blocks of 2-DE gels were acquired.
Profile of proteins was similar among the different categories of sample as most protein spots distributed inside the field of pI 4-8. ImageMaster 2-DE Platimum 5.0 software was utilized to carry out analysis for each well-matched gel dot between the comparable categories. The mean detected numbers of protein spot in each categories were as follows: 1255±43 in A45w, 1197±46 in A75w, 1198±50 in ACa, 1205±89 in ACp; 1233±30 in C45w, 1257±35 in CCa, 1244±39 in CCp; 1100±79 in D45w, 1115±61 in D105w; 1249±51 in E45w, 1179±62 in E75w, 1208±36 in E105w. Two hundred and six protein points which had double or more density and appeared in more than fifty percent gels were screened out.
One hundred sixty protein spots with different expression were dug out from the corresponding prepared Coomassie brilliant blue stain gels and one hundred forty-two spots were identified by MALDI-TOF MS. To assure the accuracy of blots sampling, each spot dug out from three blocks of gel was identified and only identical data in MS was selected. The identified differential proteins included the members of Peroxiredoxin family (PrxⅠ, PrxⅡ, PrxⅢ, and PrxⅣ), cytochrome C oxidase, glutathione peroxidaseⅠ, thioredoxin, cytochrome B5, ketohexose kinase, heterogeneous ribose nucleoprotein, phosphotriose isomerase, nicotinamide N-methyl transferase, metastasis suppressed gene NM23-H1, translation controlled tumor protein, and others.
Part 4. Verify tests on differential expression of peroxiredoxins among liver samples from tree shrews and from HCC patients
The members in peroxiredoxin family (PrxⅠ, PrxⅡ, PrxⅢand PrxⅣ), which were among the differentially expressed protein screened from Part 3 and were known had widespread biological functions concerning many other tumors, were studied further to confirm the 2-DE results that showed the expressions of PrxⅠ, PrxⅡ, and PrxⅢin HCC tissue were up-regulated greatly compared to precancerous tissue, and the expression of PrxⅣwas up-regulated prominently in HCC tissue compared to HCC-surrounding liver tissue. By Western blot and RT-PCR assays, the expression levels of PrxⅠ, PrxⅡ, PrxⅢand PrxⅣwere examined on the samples from tree shrew as well as from HCC patients.
Results confirmed that the expressions of PrxⅠ, PrxⅡ, PrxⅢ, and PrxⅣwere not only up-regulated in tree shrew HCC but also in human HCC tissue in both protein level and mRNA level, which indicated that the proteins in Prx family might play important role in hepatocarcinogenesis and could be hopefully applied as a biomarker for HCC diagnosis and/or treatment.
Conclusion
Tree shrew, a lower grade primate related closely with humankind evolutionarily, is proved suitable as animal model to research human disease because most of its laboratory values were found similar to that of human beings.
The modified 2-DE technical platform for proteomic analysis on liver tissues of tree shrews provides the foundation for research. Profiles of differentially expressed protein screened from the tissues taken at different time points during tree shrew hepatocarcinogenesis induced by AFB_1 or AFB_1+HBV suggests the progression of HCC is concerned with many factors. At least some of the proteins among the 142 identified ones might play important roles in the HCC initiation, development and/or progression. Particularly, proteins in Prx family might be applied as a biomarker for HCC diagnosis, treatment and/or prognosis-evaluation.
引文
1. Murray CJ, Lopez AD.Global health statistics,World Health Organization, Harvard School of Public Health, World Bank, 1996, 551-552.
2. Tang ZY, Ye SL, Liu YK, et al. A decade’s studies on metastasis of hepatocellular carcinoma. J Cancer Res Clin Oncol, 2004, 130(4): 187-196.
3.张思维,李连弟,鲁凤珠,等.中国1990-1992年原发性肝癌死亡调查分析.中华肿瘤杂志, 1999, 21(4): 245-249.
4. Summers J, Smolec JM, Snyder R. A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci USA, 1978, 75(9): 4533-4537.
5. Marion PL, Van Davelaar MJ, Knight SS, et al. Hepatocellurlar carcinoma in ground squirrels persistently infected with ground squirrel hepatitis virus. Proc Natl Acad Sci USA, 1986, 83(12): 4543-4546.
6. Cora L, Wild CP, Mehrotra R, et al. Contribution of aflatoxin B1 and hepatitis B virus infection in the induction of liver tumors in ducks. Cancer Res, 1990, 50(7): 2156- 2163.
7. Kim CM, Koike K, Saito J, et al. HBx gene of hepatitis B virus induces liver cancer in transgenic mice. Nature, 1992, 315(6324): 317-320.
8. Prince AM, Brotman B. Perspectives of hepatitis B studies with chimpanzees. ILAR J, 2001, 42: 85-88.
9.苏建家,严瑞琪,甘友全,等.成年树鼩实验感染人乙型肝炎病毒的研究.中华病理学杂志, 1987, 16: 103-106.
10.王荷英,刘季和,喻林冲,等.树鼩实验麻风的组织病理学观察.中华皮肤科杂志, 1996, 29(4): 258-260.
11.万新邦,庞其方,丘福禧,等.成年的中国云南树鼩对人轮状病毒易感性的实验研究.中华微生物学和免疫学杂志, 1985, 5(5): 304-307.
12.曾怀德,柴慧霞,谢扬高,等.马桑内酯引起的树鼩实验性癫痫.四川医学院学报, 1984, (2): 159-163.
13.梁真,冼苏,林健,等.树鼩糖尿病模型肝脏功能及形态学变化与TNF-α的关系.广西医科大学学报, 2003, 20(5): 639-641.
14.苏建家,李瑗,班克臣,等.树树鼩实验性肝癌发生过程p53基因的变化.中华肝脏病杂志, 2003, 11(3): 159-161.
15. Yan RQ, Su JJ,Huang DR, et al. Human hepatitis B Virus and hepatocellular carcinoma I Experimental infection of tree shrews with hepatitis B Virus. J Cancer Res clin onco, 1996, 122(5): 283-288.
16. Yan RQ, Su JJ, Huang DR, Gan YC, yang C. Huang GH. Human hepatitis B Virus and hepatocellular cainoma. II Experimental induction of hepatocllular carcinoma in tree shrews exposed to hepatitis B virus and aflatoxin B1. J Cancer Clin Oncol, 1996, 122(5): 289-295.
17.李瑗,苏建家,覃柳亮,等.乙肝病毒和黄曲霉毒素B1诱发树鼩肝癌的病理变化.临床与实验病理学杂志, 2000, 16(3): 224-227.
18. Li Y, Su JJ, Qi LL,et al. Synergistic effect of hepatitis B Virus and aflatoxin B1 in hepatocarcinogenesis in tree shrews. Ann Acad Med Singapore, 1999, 28(1): 67-71.
19.李瑗,苏建家,曹骥,等.不同因素诱发的树鼩肝癌组织的基因表达差异.癌症, 2003, 22(10): 1018-1022.
20.何琼琼,程瑞雪,陈主初.蛋白质组学在肝癌研究中的应用.国外医学肿瘤学分册, 2005, 32(10): 782-784.
21.许克新,王云川.双向电泳及质谱分析与蛋白质组研究.第四军医大学学报, 2002, 23(22): 2017-2022.
22. O’Farrel PH. High resolution Two-dimensional electrophoresis of proteins. J Bilo chem, 1975, 20:4007.
1.郭鹞.人类疾病的动物模型.第一版.北京:人民卫生出版社, 1990: 1-6.
2.彭燕章,叶智彰,邹如金,等.树鼩生物学.昆明:云南科技出版社, 1991: 1-4.
3.严瑞琪,苏建家,陈志英,等.人乙型肝炎病毒实验感染成年树鼩的初步研究.广西医学院学报, 1984, 1(1): 10-15.
4.苏建家.实验性肝细胞癌研究状况.医学文选, 2001, 20(1): 1-3.
5. Brewer NR. Personalities in the early history of laboratory animal science and medicine.LabAnisci, 1980, 30(4): 741-758.
6. Crafton TS. The founding and early history of the national society of medical research. Lab Anisci, 1980, 30(40): 760-764.
7. Park US, SuJJ,Ban KC, et al. Mutations in the p53 tumoe suppressor gene in tree shrew hepatocellular carcinoma associate with hepatitis Bvirus infection and intake of aflatoxin B1.Gene, 2000, 251(1): 73-80.
8.李瑗,苏建家,曹骥,等.用cDNA阵列技术研究黄曲霉素B1诱发树鼩肝癌过程中的基因变化.中华肝脏病杂志, 2003, 11(2): 96-99.
9.詹美云,刘崇柏,李成明,等.甲型肝炎病毒感染树鼩的初步研究.中国医学科学院学报, 1981, 3(3): 148-152.
10.王凤水,陈红松,王宇,等.树鼩感染人乙肝病毒动物模型研究中的非特异性反应.北京医科大学学报, 1996, 28(5): 340-342.
11.刘志,毛青,王宇明,等.丙型肝炎病毒感染成年树鼩的实验研究.第三军医大学学报, 1998, 20(6): 472-475.
12.李奇芬,丁明权,王洪,等.成年树鼩实验感染丁型肝炎病毒的研究.第三军医大学学报, 1994, 16(4): 234.
13.李奇芬,毛青,吴纯清,等.树鼩实验性丁型肝炎的病理观察.中华肝脏病杂志, 1996, 4(4): 210-213.
14.李奇芬,毛青,吴纯清,等.树鼩传代感染HDV后肝脏内抗原表达及病理变化.世界华人消化杂志, 1999, 7(5): 378-381.
15.万新邦,庞其方,丘福禧,等.成年的中国云南树鼩对人轮状病毒易感性的实验研究.中华微生物学和免疫学杂志, 1985, 5(5): 304-307.
16. Darai G, Zoller L, Matz B, et al. Experimental infection and the state of viral latency of adult Tupaia with herpes simplex virus type 1 and 2 and infection of juvenile Tupaia with temperature-sensitive mutants of HSV type 2. Arch Virol, 1980, 65(3-4): 311-318.
17.戴长柏.人单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染树鼩的研究.医学生物学研究, 1984, 1: 62-69.
18.朱宇同,王维祥,曾子安,等.流感病毒感染成年树鼩的研究.上海实验动物科学, 1986, 6(6): 20-23.
19.刘丽华,李正平,俞永新.树鼩对登革热病毒敏感性的研究.中华微生物学和免疫学杂志, 1985, 5(5): 329-330.
20.侯宗柳,白登云,黄文丽,等.树鼩对森林脑炎病毒的敏感性及发病机理的研究.中国病毒学, 1991, 6(4): 298-303.
21.鲍铭晖,钱小榆,张海林,等.树鼩感染基孔肯雅病毒的病理学研究.云南医药, 1992, 13 (3): 180-182.
22.张海林,米竹青,施华芳,等.树鼩实验感染后血清中抗基孔肯雅病毒IgM和IgG抗体检查.地方病通报, 1997, 12(2): 26-27.
23. McBrien NA, Cornell LM, Gentle A. Structural and ultrastructural changes to the sclera in a mammalian model of high myopia. Invest Ophthalmol Vis Sci, 2001, 42(10): 2179-2187.
24. Gentle A, Liu Y, Martin JE, Conti GL, et al. Collagen gene expression and the altered accumulation of scleral collagen during the development of high myopia. J Biol Chem, 2003, 278(19): 16587-16594.
25. Fuchs E, Flugge G.Psychosocial stress induces molecular and structural alterations in the brain - How animal experiments help to understand pathomechanisms of depressive illnesses. Psychosom Med Psychother, 2001, 47(1): 80-97.
26. Fuchs E. Social stress in tree shrews as an animal model of depression: an example of a behavioral model of a CNS disorder. CNS Spectr, 2005, 10(3): 182-189.
27.苏建家,李瑗,班克臣,等.树鼩实验性肝癌发生过程p53基因的变化.中华肝脏病杂志, 2003, 11(3): 159-161.
28. Ishiko S, Yoshida A, Mori F,et al. Early ocular changes in a tree shrew model of diabetes.Nippon Ganka Gakkai Zasshi, 1997, 101(1): 19-23.
29. Steinhausen M, Thederan H, Nolinski D,et al. Acute renal failure in Tupaia belangeri and rats. Bibl Anat, 1979, (18): 90-92.
30.曾怀德,柴慧霞,谢扬高,等.马桑内酯引起的树鼩实验性癫痫.四川医学院学报, 1984, 2: 159-163.
31. Schwaier A. Tupaias (tree shrews)--a new animal model for gallstone research; first observations of gallstones. Res Exp Med (Berl), 1979, 176(1):15-24.
32.谈跃,王子灿,李麟仙.树鼩局部脑缺血模型的研究.昆明医学院学报, 1994, 15(3): 17-21.
33. Braun H, Kloft W. Hamatologische untersuchngen am spizhornchcn Tupata lana.Tupaiidae Prosimiae, 1965, 21: 663-664.
34. Hunt RD, Chalifoux L. The hemogram of the tree shrew. Folia Priimat, 1967, 7: 34-36.
35.邹如金,代伟,昆龙,等.树鼩血相的研究.动物学研究, 1983, 4(3): 291-293.
36.李瑗, Karen Baumgartner, Denise MacMillan,等.树鼩的人工哺育.上海实验动物科学, 2001, 21(2): 70-74.
1.许克新,王云川.双向电泳及质谱分析与蛋白质组研究.第四军医大学学报, 2002, 23(22): 2017-2022.
2.郭尧君.SDS电泳技术的实验考虑及最新进展.生物化学与生物物理学进展, 1991, 18: 32-35.
3. O’Farrel PH. High resolution Two-dimensional electrophoresis of proteins.J Bilo chem, 1975, 20: 4007.
4. Cordwell SJ, Nouwens AS, Verrills NM,et al. Subproteomics based upon protein cellular location and relative solubilities in conjunction with composite two-dimensional dldtrophoredid gels. Electrophoresis, 2000, 21:1094-1103.
5. Gorg A, Obermaier C, Boguth G,et al. The current state of two-dimensional eletrophoresis with immobilized pH gradients.Eletrophoresis, 2000, 32: 1037 -1053.
6.朱雪琼,石一夏,陈怀增,等.子宫肌瘤蛋白质组研究中二维电泳方法的建立.浙江大学学报(医学版), 2004, 6(33): 550-552.
7.郭尧君.载体两性电解质pH梯度等电聚焦.见:郭尧君,主编.蛋白质电泳实验技术.第一版.北京:科学出版社出版, 1999, 161-210.
8. Fountoulakis M, Takacs B. Effect of strong detergents and chaotropes on the Deiection of proyeins in two-dimensional gels.Eletrophoresis, 2001, 22(9): 1593-1602.
9. Molloy MP. Two-dimensional eletrophoresis of membrane proteins using immobilizsd pH gradients.Anal Biochem, 2000, 280(1): 1-10.
10. Rabillooud T, Adessi C, Giraudel A, et al. Improcement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients. Eletrophoresis, 1997, 18: 307-316.
11. Herbert BR, Molloy MP, Gooley AA, et al. Improved protein solubility in two-dimensional electrophoresis using tributyl phosphine as reducing agent.Electrophoresis, 1998, 19: 845-851.
12. Gorg A,Obermaier C,Boguth G,et al.The current state of two-dimensional electrophoresis with immobilized pH gradients.Electrophoresis, 2000, 21(6): 1037-1053.
13. Qian xiao-hong.Proteomics: from Protein Sequence to Function. Beijing: Scientific Publisher, 2002, 57-58.
14. Yan JX, Wait R, Berkelman T, et al. A moditidf silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ioni-zation and electrospray ionization-mass spectrometry. Electrophresis, 2000, 21(17): 3666-3667.
15.Brush M. Dye haed: protein gel staining products.Scientific, 1998, 12: 16-22.
16.Neuhoff V, Arold N, Taube D, Ehrhardt W. Improved staining of proteins in polyacrylamide gels including isorlrctric focusing Gels with clear background at nanogram sensitivity using Coomassir Brilliant Blue G-250 and R-250.Electrophoresis, 1988, 9(6): 255-262.
17.Neuhoff V, Stamm R, Pardowitz I, et al. Essential problems in quantification og preteins following cloolidal staining with coomassie brilliant blue dyes in polyacrylamide gels, and rheir solution.Electrophoresis, 1990, 11(2): 101- 117.
1.王树声,苏建家,张华远,等.树鼩、熊猴感染人乙型肝炎病毒血清指症的研究.中华实验和临床病毒学杂志, 2003, 17(1): 21-24.
2. Kim J, Kim SH, Lee SU, et al. Proteome analysis of human liver tumor tissue by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of disease-related proteins. Electrophoresis, 2002, 23(24): 4142-4156.
3. Cha C, Dematteo RP. Molecular mechanisms in hepatocellular carcinoma development. Best Pract Res Clin Gastroenterol, 2005, 19(1): 25-37
4. Ming L, Thorgeirsson SS, Gail MH, et a1. Dominant role of hepatitis B virusand cofactor role of aflatoxin in hepatocarcinogenesis in qidong China. Hepatotology, 2002, 36(5): 1214-1220
5. Groopman JD, Wang JS, Scholl P. Molecular biomarkers for aflatoxins:from adducts to gene mutations to human liver cancer.Can J Physiol Pharmacol, 1996, 4(2): 203-209.
6.苏建家,李瑗,班克臣,等.动态研究HBV和AFB1诱发树鼩肝细胞癌过程中一些基因的表达,广西医科大学报, 2001, 18(2): 151-154.
7. Garrido C, Galluzzi L, Brunet M, et al. Mechanisms of cytochrome c release from mitochondria.Cell Death and Differentiation , 2006,5: 1–11.
8. Yanagawa T, Omura K, Harada H, et a1.Peroxiredoxin I expression in tongue squamous cell carcinomas as involved in tumor recurrence. Int J Oral Maxillofac Surg, 2005, 34(8): 915-920.
9. Shen J, Person MD, Zhu J,et al. Protein expression profiles in pancreatitis as detected by two-dimensional gel electrophoresis and mass spectrometry. Cancer Res, 2004, 64(24): 9018-9026.
10. Kim HJ, Chae HZ, Kim YJ, et a1.Preferential elevation of Prx I and Trx expression in lung cancer cells following hypoxia and in human lung cancer tissues.Cell Biol Toxicol, 2003, 19(5): 285-298.
11. Crowley-Weber CL, Payne CM, Gleason-Guzman M, et a1.Development and molecular characterization of HCT-116 cell lines resistant to the tumor promoter and multiple stress-inducer deoxycholate. Carcinogenesis,2002, 23(12): 2063-2080.
12. Choi JH,Kim TN,Kim S,et a1.Overexpression of mitochondrial thioredoxin reductase and peroxiredoxin III in hepatocellular carcinomas. Anticancer Res, 2002, 22(6A): 3331-3335.
13. Jang JS, Cho HY, Lee YJ, et al. The differential proteome profile of stomach cancer: identification of the biomarker candidates.Oncol Res, 2004, 14(10): 491-499.
14. Yong-Zheng Wu, Yefim Manevich, James L, et al. Interaction of SurfactantProtein A with Peroxiredoxin 6 Regulates Phospholipase A2 Activity.Biol. Chem, 2006, 11(281): 7515-7525.
15. Miyazawa1 T, Kubo1 E, Fatma N, et al. Peroxiredoxin6–Deficient Mouse Lens Epithelial Cells Exhibit an Increased Susceptibility to Hypergylycemia Induced Apoptosis. Invest Ophthalmol Vis Sci, 2005(46): 3859.
16. Xuesong Ouyang, Theodore L, DeWeese, et al. Loss-of-Function of Nkx3.1 Promotes Increased Oxidative Damage in Prostate Carcinogenesis. Cancer Research, 2005(65): 6773-6779.
17.周海君,刘银坤,崔杰峰,等.人肝癌细胞系的糖蛋白质组学研究.生物化学与生物物理进展, 2006, 33(1): 59-64.
18. Varesco L, Caligo MA, Simi P, et al. The NM23 gene maps to human chromosome band 17q22 and shows a restriction fragment length polymorphism with BgⅠ,Ⅱ.Genes Chromosom Cancer, 1992, 4(1):84.
19. Urano T, Furuhawa K, Sbihu H. Expression of NM23/NDP kinase proteins on the cell surface.Oncogene, 1993, 8: 1371.
20. Leone A, Flatow U, King CR, et al. Reduced tumor incidence, metastatic potential and cytokine responsiveness of NM23-transfected melanoma cell.Cell, 1991, 65(1): 25-35.
21. Ennessy C, Henry JA, May FEB, et al. Expression of the antimetastatic gene NM23 in human breast cancer. J Natl Cancer Inst, 1991, 83: 281.
22. Irayama R, Sawai S, Takagi Y, et al. Positive relationship between expression of anti-metastatic factor (NM23 gene product or nucleotide diphosphate kinase) and good prognosis in human breast cancer. J NatI Cancer Inst, 1991, 83: 1249.
23. Nakayama H,Yasui W,Yokozaki H,et al. Reduced expression of NM23 is associated with metastasis of human gastric carcinomas. Jpn J Cancer Res, 1993, 84(2): 184-190.
24. Müller W, Schneiders A, Hommel G, et al. Expression of NM23 in gastric carcinoma:association with tumor progression and poor prognosis1. Cancer,1998, 83(12): 2481.
25. Yamaguchi A, Urano T, Fushida S, et al. Inverse association of NM23-H1 expression by colorectal cancer with liver metastasis.Br J Cancer 1993, 68(5): 1020-1024.
26. Nakayama T, Ohtsuru A,Nakao K, et al. Expression in human hepatocellular carcinoma of nucleotide diphosphate kinase,a homologue of the NM23 gene product. J Natl Cancer Inst, 1992, 84:1349.
27. Rosengard AM, Krutzsch HC, Sbearn A, et al. Reduced NM23/and protein in tumor metastasis and abberant drosphila development.Nature, 1989, 342: 177.
28.盛红光,张敏,姜言明,等.肿瘤血管形成和转移抑制基因NM 23与肝细胞癌侵袭性关系的研究.中华普通外科, 2004, 19(3): 180.
29. Lombardi D, Lacombe ML, Paggi MG. NM23:unraveling its biological function in cell differentiation.J Cell Physio, 2000, 82(2): 144.
30. Caligo MA, Cipollini G, Fiore L, et al. NM23 gene expression correlates with cell growth rate and sphate. Int J Cancer, 1995, 60(6): 837.
31. Kantor JD, McCormick B, Steeg PS, et al. Inhibition of cell mortality after NM23 transfection of human and murine tumor cell.Cancer Res, 1993, 53 (9): 1971.
32.张豪,孙桂菊.乙肝病毒和化学致癌物致肝细胞癌变的研究进展.肿瘤, 2004, 24(5): 509-511.
33.李瑗,苏建家,曹骥,等.用cDNA阵列技术研究黄曲霉毒素B1诱发树鼩肝癌形成过程中的基因变化.中华肝脏病杂志, 2003, 11(2): 96-99.
34. Allred DC, Avis I, et a1.Diferentialexpressionofthe early lung can cer detection marker, heterogeneous nuclear ribonucleopmtein-A2/B1 innormal breast abreastand neoplastic breastcancer.Breast CancerRes,2001, 66: 217-224.
35. Wu S, Sato M, Endo C, Sakurada A, et a1. hnRNP B1 protein may be a possible prognostic factor in squamous cell carcinoma of the lung. LungCancer, 2003, 41(2): 179-186.
36. Satoh H, Kamma H, Ishikawa H, et a1.Expression of hnRNP A2/B1 proteins in human cancer cell lines. Int J Oncol, 2000, 16(4): 763-767.
37. Ushigome M, Ubagai T, Fukuda H, et a1.Up-regulation of hnRNP A1 gene in sporadic human colorectal cancers. Int J Oncol, 2005, 26(3): 635-640.
38. Christian K, Lang M, et a1. Interaction of heterogeneous nuclear ribonucleoprotein A1 with cytochrome P450 CYP2A6 mRNA: implications for post-transcriptional regulation of the CYP2A6 gene. Mol Pharmacol, 2004, 65(6): 1405-1414.
39. Kamma H, FujimotoM, FujiwaraM, et a1. Interactionofhn-RNP A2/B1 isoforms with telomefic ssDNA and the invitro function.Biochem Biophy Res Comm, 2001, 280: 625-630.
40. Iwanaga K, Sueoka N, Sato A,et al.Heterogeneous nuclear ribonucleoprotein B1 protein impairs DNA repair mediated through the inhibition of DNA-dependent protein kinase activity. Biochem Biophys Res Commun, 2005, 333(3): 888-895.
41. Jang JS, Cho HY, Lee YJ, et al. The differential proteome profile of stomach cancer: identification of the biomarker candidates. Oncol Res, 2004, 14 (10): 491-499.
42. Gehrmann ML,Fenselau C,Hathout Y. Highly altered protein expression profile in the adriamycin resistant MCF-7 cell line. J Proteome Res, 2004, 3(3): 403-409.
43. Jimin Xu, Marco Capezzone, Xiao Xu, et al. Activation of Nicotinamide N-Methyltransferase Gene Promoter by Hepatocyte Nuclear Factor-1? in Human Papillary Thyroid Cancer Cells. Molecular Endocrinology, 2005, 19(2): 527-539
44. Narimatsu T,Tamori A,Koh N,et a1.p16 promoter hypermethylation in human hepatocellular carcinoma with or without hepatitis virus infection. Intervirology, 2004, 47(1): 26-31.
45.吴曼,薛开先,孙开来,等.肿瘤遗传学.北京:科学出版社.2004: 3-26.
46. Pang R, Yuen J, Yuen MF, et a1. PIN 1 overexpression and beta-catenin gene mutations are distinct oneogenie events in human hepatocellular carcinoma. Oneogene, 2004, 23 (23): 4182-4186.
47. Zhong S,Yeo W,Tang MW, et a1.Intensive hyperm ethylation of the CpG island of Ras association domain family lA in hepatitis B virus-associated hepatocellular carcinomas.Clin Cancer Res, 2003, 9(9): 3376-3382.
48. Heilbronn R, Sehlehofer JR, Yalkinoglu AO, et al. Selective DNA- amplification induced by carcinogens (initiators): evidence for a role of proteases and DNA polymerase alpha. Int J Cancer, 1985, 36(1): 85-91.
49. Zhang YJ, Ahsan H, Chen Y, et a1. High frequency of promoter hyperm ethylation of RASSF1A and p16 and its relationship to aflatoxin B1-DNA adduct levels in human hepatocellular carcinoma. Mol Carcinog, 2002, 35(2): 85-92.
50. Sohn S,Jaitovitch-Groisman I,Benlimame N,et a1.Retroviral expression of the hepatitis B virus X gene promotes liver cell susceptibility to carcinogen-induced site specific mutagenesis.Mutat Res,2000,460(1):17.
51. Livezey KW, Negorev D, Simon D, et a1.Increased chromosomal alterations and micronuclei formation in human hepatoma HepG2 cells transfected with the hepatitis B virus HBX gene. Mutat Res, 2002, 505(1-2): 63.
52. Capovilla A, Arbuthnot P. Hepatitis B virus X protein does not influence essential steps of nucleotide excision repair effected by human liver extracts. Biochem Biophys Res Commun, 2003, 312(3): 806.
53. Wild CP, Shrestha SM, Anwar WA, er al. Field studies of aflatoxin exposure, metabolism and induction of genetic alterations in relation to HBV infection and hepatocellular carcinoma in The Gambia and Thailand. Toxicol Lett, 1992, 64-65: 455-461.
54. Bontron S, Lin-Marq, Strubin M, et a1. Hepatitis B virus X proteinassociated with UV-DDB1 induces cell death in the nucleus and is functionally antagonized by UV-DDB2.J Biol Chem,2002,277(41):3884
55. liams CS,Dubois RN. Prostaglandin endoperoxide synthase: Why two isoforms .Am J Physiol, 1996, 270(1): 393.
56. Sadrzadeh SM, Nanji AA. The 212aminosteroid 162desmethyl tirilazad mesylate prevents necroinflammatory changes in experimental alcoholic liver disease. J Pharmacol Exp Ther, 1998, 284(1): 406.
57. Nanji AA, Miao L, Thomas P, et al. Enhanced cyclooxygenase gene expression in alcoholic liver disease in the rat. Gastroenterology, 1997, 112 (3): 943.
58. Nieto N, Greenwel P, Friedman SL, et al. Ethanol and arachidonic acid increase alpha 2(I)collagen expression in rat hepatic stellate cells overexpressing cytochrome P450 2El; role of H2O2 and cyclooxygenase-2.J Biol Chem, 2000, 275(26): 20136-20145.
59. Weiss P, Mouallem M, Bruck R, et al. Nimesulide2induced hepatitis and acute liver failure.Isr Med Assoc J , 1999, 1(2): 89.
60. Shiota G, Okubo M, Noumi T, et al. Cyclooxygenase expression in hepatocel lularcarcionoma.Hepatogastroenterology, 1999, 46(25): 407.
61. Koga H, Sakisaka S, Ohishi M, et al. Expression of COX-2 in human hepatocellular carcinoma : relevance to tumor dedifferentation . Hepatology, 1999, 29(3): 688
62. Choi JH, Kim TN, Kim S, et a1. Overexpression of mitochondrial thioredoxin reductase and peroxiredoxinⅢin hepatocellular carcinomas. Anticancer Res, 2002, 22(6A): 3331-3335.
63. Haapasalo H, Kylaniemi M, Paunul N, et a1. Expression of antioxi-dant enzymes in astrocytic brain tumors. Brain Pathol, 2003, 13(2): 155-164.
64. Lincoln DT, Ali Emadi EM, Tonissen KF, et a1.The thioredoxin -thioredoxin reductase system: over-expression in human cancer. Anticancer Res, 2003, 23(3B): 2425-2433.
65. Gromer S,Gross JH.Methylseleninate is a substrate rather than an inhibitor of mammalian thioredoxin reductase.Implications for the antitumor effects of selenium. J Biol Chem, 2002, 277(12): 9701-9706.
66. Edamoto Y,Hara A,Biernat W,et a1. Alterations of RB1, p53 and W nt pathways in hepatocellular carcinomas associated with hepatitis C,hepatitis B and alcohol liver cirrhosis. Int J Cancer, 2003, 106(3): 334-341.
67. Szabo E,Paska C,Kaposi Novak P,et a1.Similarities and difierences in hepatitis B an d C virus induced hepatocarcinogenesis.Pathol Oncol Res,2004, 10(1): 5-11. .
1. Rhee SG, Kang SW, Chang TS, et al. peroxiredoxin, a novel family of peroxi -dases. IUBMB Life, 2001, 52(1-2): 35-41.
2. Fujii J, Ikeda Y. Advances in our understanding of peroxiredoxin, a multi- functional,mammalian redox protein. Redox Rep, 2002, 7(3): 123-130.
3. Tien Nguyen-nhu N, Knoops B. Mitochondrial and cytosolic expression of human peroxiredoxin 5 in Saccharomyces cerevisiae protect yeast cells from oxidative stress induced by paraquat. FEBS Lett, 2003, 544: 148-152.
4. ChangT S,Jeong W,Choi SY,et al.Regulation of peroxiredoxin I activity by Cdc2-mediated phosphorylation. J Biol Chem, 2002, 277(28):25370- 25376.
5. Wood Z A, Schroder E, Robin Harris J, et a1. Structure, mechanisnl and regulation of peroxiredoxins.Trends Biochem Sc1, 2003, 28 (1): 32-40
6. Fujii J, Ikeda Y. Advances in our understanding of peroxiredoxin, a multi -functional, mammalian redox protein. Redox Rep, 2002, 7(3):123-130.
7. Henkle-Duhrsen K, Kanapkotter A.Antioxidant enzvyme families in parasitic nematodes. Mol Biochem Parasitol, 200l, 114(2): l29-l42.
8. Rabilloud T, Heller M, Gasnier F, et a1. Proteonfics analvsis of cellular response to oxidative stress.Evidence for in vivo overoxidation of peroxire -doxins at their active site. J Biol Chem, 2002, 277(22): l9396-1940l.
9. Zhou Y,Kok K H,Chun A C,et al.Mouse peroxiredoxin V is a:thioredoxin peroxidase that inhibits p53-induced apoptosis.Biochem Biophys Res Commun, 2000, 268(3): 921-927.
10. Shan H,Butterfield L H,Chiu R,et al.Cloning and sequeuce analysis of candidate human natural killer-enhancing factor genes. Immunogenetics, 1994, 40(2): 129-134.
11.章波,向渝梅,白云.抗氧化蛋白Peroxiredoxin家族研究进展.生理科学进展, 2004, 35(4): 352-355.
12.周海君,刘银坤,崔杰峰,等.人肝癌细胞系的糖蛋白质组学研究.生物化学与生物物理进展, 2006, 33(1): 59-64.
13. Crowley-Weber CL, Payne CM, Gleason-Guzman M, et a1.Development and molecular characterization of HCT-116 cell lines resistant to the tumor promoter an d multiple stress-inducer,deoxycholate.Carcinogenesis, 2002, 23: 2063-2080.
14. Kim HJ, Chae HZ, Kim YJ, et a1. Preferential elevation of Prx I and Trx expression in lung cancer cells following hypoxia and in human lung cancer tissues.Cell Biol Toxicol, 2003, 19(5): 285-298.
15. Jang JS, Cho HY, Lee YJ, et al. The differential proteome profile of stomach cancer: identification of the biomarker candidates.Oncol Res, 2004, 14(10): 491-499.
16. Chang TS, Jeong W, Choi SY,et al. Regulation ofPeroxiredoxin I Activity by Cdc2-mediated Phosphorylation .J Biol Chem, 2002, 277(28): 25370-25376.
17. Gisin J, Perren A, Bawohl M, Rare allelic imbalances, but no mutations of the PRDX1 gene in human hepatocellular carcinomas. Journal of Clinical Pathology, 2005, 58(11): 1229- 1231.
18. Choi MH, Lee IK, Kim GW, et al. Regulation of PDGF signalling and vascular remodelling by peroxiredoxin II. Nature, 2005, 435(7040): 347-353.
19. Kim H, Lee TH, Park ES, et al. Role of peroxiredoxins in regulating intra -cellular hydrofen peroxide and hydrogen peroxide-induced apoptosis in thyroid cell. J Biol Chen, 2000, 275(24): 18266-18270.
20.王世鄂,黄威权,付建芳等. Peroxiredoxin II在小鼠卵巢中卵泡、GV卵、MII卵和植入前胚的表达和定位.解剖学报, 2004, 35(6): 647-650.
21. Araki M.Nanri H.Ejima K,et al.Antioxidant function of the mitochondrial protein SP-22 in the cardiovascular systen1. J Biol Chem, l999, 274(4): 227l -2278.
22. Shih S F,Wu Y H,Hung CH,et a1.Abrin triggers cell death by inactivating a thiol-specific antioxidant protein. J Biol Chem, 200l, 276 (24): 2l870-2l877.
23. Masaki M,Ikeda A,Shiraki E,et al.Mixed lineage kinase LZK and anti -oxidant protein-l activate NF-kappaB synergistically.Eur J Biochem, 2003, 270(1): 76-83.
24. Jasehke A.Mi H.Fropschug M.Human T cell cyelophilin 18 binds to thiol- specific antioxidant protein Aopl and stimulates its activity. J Mol Biol, 1998, 277(4): 763-769.
25.冯艳,刘东青,王震,等.心肌特异性新蛋白激酶p93和peroxiredoxin 3的相互作用.生物化学与生物物理进展, 2004, 31(8): 688-691.
26. Larisa Nonn1, Margareta Berggren, et al. Increased Expression of Mitochondrial Peroxiredoxin-3 (Thioredoxin Peroxidase-2) Protects Cancer Cells Against Hypoxia and Drug-Induced Hydrogen Peroxide-Dependent Apoptosis1. Molecular Cancer Research, 2003, 1: 682-689.
27. Tong-Shin Chang, Chun-Seok Cho, Sunjoo Park, et al. Peroxiredoxin III, a Mitochondrion-specific Peroxidase, Regulates Apoptotic Signaling byMitochondria. J Biol Chem, 2004, 279(40): 41975-41984.
28. Kamma H,FujimotoM,FujiwaraM,et a1.Interactionofhn-RNP A2/B1 isoforms with telomefic ssDNA and the invitro function.Biochem Biophy Res Comm, 2001, 280:625-630.
29. Miyazawa1 T, Kubo1 E,Fatmay N,et al.Peroxiredoxin6–Deficient Mouse Lens Epithelial Cells Exhibit an Increased Susceptibility to Hypergylycemia Induced Apoptosis. Invest Ophthalmol Vis Sci, 2005(46): 3859.
30. Yan Wang, Shelley A. Phelan, Yefim Manevich, Sheldon I. Transgenic Mice Overexpressing Peroxiredoxin 6 Show Increased Resistance to Lung Injury in Hyperoxia. American Journal of Respiratory Cell and Molecular Biology, 2006(34): 481-486.
1詹美云,刘崇柏,李成明,等.甲型肝炎病毒感染树鼩的初步研究.中国医学科学院学报, 1981, 3 (3): 148-152.
2严瑞琪,苏建家,陈志英,等.人乙型肝炎病毒实验感染成年树鼩的初步研究.广西医学院学报, 1984, 1(1): 10-15.
3苏建家,严瑞琪,甘友全,等.成年树鼩实验感染人乙型肝炎病毒的研究.中华病理学杂志, 1987, 16(2): 103-106.
4王凤水,陈红松,王宇,等.树鼩感染人乙肝病毒动物模型研究中的非特异性反应.北京医科大学学报, 1996, 28(5): 340-342.
5王海平,姚志强,周永兴,等.树鼩实验感染乙型肝炎病毒的再评价.中华肝脏病杂志, 1998, 6(2): 96-97.
6苏建家,李瑗,王树声,等.树鼩对人乙型肝炎病毒易感性的验证.广西医学, 2000, 7(4): 298-301.
7王树声,苏建家,张华远,等.树鼩、熊猴感染人乙型肝炎病毒血清指征的研究.中华实验和临床病毒学杂志, 2003, 17(1): 21-24.
8王海平,周永兴,姚志强,等.成年树鼩实验感染丙型肝炎病毒的初步研究.第四军医大学学报, 1997, 18(4): 375-376.
9刘志,毛青,王宇明,等.丙型肝炎病毒感染成年树鼩的实验研究.第三军医大学学报,1998,20(6): 472-475.
10 Xie ZC, Riezu-Boj JI, Lasarte JJ, et al. Transmission of hepatits C virus infection to tree shrews. Virology, 1998, 244(2): 513-520.
11谢志春,高伟志,苏洁寒,等.树鼩对丙型肝炎病毒的易感性研究.广西医科大学学报, 2000, 17(3): 347-350.
12 Zhao X, Tang ZY, Klumpp B,et al. Primary hepatocytes of Tupaia belangeri as a potential model for hepatitis C virus infection. J Clin Invest. 2002,109(2):221-32.
13李奇芬,丁明权,王洪,等.成年树鼩实验感染丁型肝炎病毒的研究.第三军医大学学报, 1994, 16(4): 234.
14李奇芬,毛青,吴纯清,等.树鼩实验性丁型肝炎的病理观察.中华肝脏病杂志, 1996, 4(4): 210-213.
15李奇芬.毛青.吴纯清.等.树鼩传代感染HDV后肝脏内抗原表达及病理变化.世界华人消化杂志, 1999, 7(5): 378-381.
16谢志春,周元平,吴继周,等. TTV感染成年树鼩的初步研究.广西预防医学, 2000, 6(3): 133-135.
17万新邦,刘玑昌,胥爱源,等.用人轮状病毒感染成年树鼩的实验研究.中华医学杂志, 1982, 62 (8): 461-465.
18万新邦.中国云南成年树鼩对人轮状病毒的易感性的实验研究.医学生物学研究, 1985, 1: 1-10
19万新邦,庞其方,丘福禧,等.成年的中国云南树鼩对人轮状病毒易感性的实验研究.中华微生物学和免疫学杂志, 1985, 5(5): 304-307.
20 Darai G, Zoller L, Matz B,et al.Experimental infection and the state of viral latency of adult Tupaia with herpes simplex virus type 1 and 2 and infection of juvenile Tupaia with temperature-sensitive mutants of HSV type 2.Arch Virol, 1980, 65(3-4): 311-318.
21徐维明.人单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染树鼩的超微结构研究.医学生物学研究, 1984, (1): 73-77.
22戴长柏.人单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染树鼩的研究.医学生物学研究, 1984,(1): 62-69.
23朱宇同,王维祥,曾子安,等.流感病毒感染成年树鼩的研究.上海实验动物科学, 1986, 6(6): 20-23.
24刘丽华,李正平,俞永新.树鼩对登革热病毒敏感性的研究.中华微生物学和免疫学杂志, 1985, 5(5): 329-330.
25侯宗柳,白登云,黄文丽,等.树鼩对森林脑炎病毒的敏感性及发病机理的研究.中国病毒学, 1991, 6(4): 298-303.
26鲍铭晖,钱小榆,张海林,等.树鼩感染基孔肯雅病毒的病理学研究.云南医药, 1992, 3: 180-182.
27张海林,米竹青,施华芳,等.树鼩实验感染后血清中抗基孔肯雅病毒IgM和IgG抗体检查.地方病通报, 1997, 12(2): 26-27.
28王荷英,叶顺章,刘季和,等.麻风菌接种树鼩的研究.中国麻风杂志, 1989, 5(3): 137-139.
29薛文昌,韩永高,陈剑英.等.麻风杆菌已经能在树鼩足垫中生长:麻风杆菌又一个新的动物模型.皮肤病与性病, 1990, 12(1): 1-5.
30王荷英,刘季和,喻林冲,等.树鼩实验麻风的组织病理学观察.中华皮肤科杂志, 1996, 29(4): 258-260.
31王志光,朱杰玲,武云,等.树鼩人工感染马锥虫的研究.上海实验动物科学, 1987, 7(2):79.
32 Ishiko S, Yoshida A, Mori F,et al. Early ocular changes in a tree shrew model of diabetes. Nippon Ganka Gakkai Zasshi, 1997, 101(1): 19-23.
33彭葆坤,周宁,唐哲,等.实验性糖尿病树鼩四周心肌超微结构改变和心脏血液动力学变化.中华临床医药杂志, 2004, 5(4): 8-11.
34梁真,冼苏,林健,等.树鼩糖尿病模型肝脏功能及形态学变化与TNF-α的关系.广西医科大学学报, 2003, 20(5): 639-641.
35谈跃,王子灿,李麟仙.树鼩局部脑缺血模型的研究.昆明医学院学报, 1994, 15(3): 17-21.
36王子灿,李麟仙,谈跃,等.树鼩局部脑缺血模型病理生理特点及实验治疗的研究.中国病理生理杂志, 1997,13(5): 492.
37 Steinhausen M, Thederan H, Nolinski D,et al. Acute renal failure in Tupaia belangeri and rats. Bibl Anat. 1979,(18):90-2.
38曾怀德,柴慧霞,谢扬高,等.马桑内酯引起的树鼩实验性癫痫.四川医学院学报, 1984, 2 : 159-163.
39 Schwaier A. Tupaias (tree shrews)--a new animal model for gallstone research; first observations of gallstones. Res Exp Med (Berl), 1979, 176(1):15-24.
40佘铬鹏,夏人仪,王宗立,等.树鼩高密度脂蛋白(HDL)抗动脉粥样硬化的研究:一种新的动物模型.医学研究通讯, 1985, 3 : 89-90.
41 Zadnik K,Mutti DO.How applicable are animal myopia models to human juvenile onset myopia?Vision Res, 1995, 35(9): 1283-1288.
42 Edwards MH.Animal models of myopia.A review.Acta Ophthalmol Scand, 1996, 74(3): 213-219.
43 McBrien NA, Cornell LM, Gentle A. Structural and ultrastructural changes to the sclera in a mammalian model of high myopia.. Invest Ophthalmol Vis Sci, 2001, 42(10): 2179-2187.
44 Gentle A, Liu Y, Martin JE, Conti GL,et al. Collagen gene expression and the altered accumulation of scleral collagen during the development of high myopia. J Biol Chem, 2003, 278(19): 16587-16594.
45 Fuchs E, Flugge G. Psychosocial stress induces molecular and structural alterations in the brain - How animal experiments help to understand pathomechanisms of depressive illnesses Psychosom Med Psychother. 2001, 47(1): 80-97.
46 Czeh B, Michaelis T, Watanabe T,et al. Stress-induced changes in cerebral metabolites, hippocampal volume, and cell proliferation are prevented by antidepressant treatment with tianeptine. Proc Natl ACad Sci U S A, 2001 , 98(22): 12796-12801.
47 Fuchs E, Flugge G. Social stress in tree shrews: effects on physiology, brain function, and behavior of subordinate individuals. Pharmacol Biochem Behav, 2002 ,73(1):247-258.
48 Fuchs E. Social stress in tree shrews as an animal model of depression: an example of a behavioral model of a CNS disorder. CNS Spectr. 2005 , 10(3): 182-189.
49严瑞琪,苏建家,黄定瑞,等.人乙型肝炎病毒和黄曲霉毒素B1诱发树鼩原发性肝癌的研究.中华病理学杂志, 1989, 18(1): 19-22.
50严瑞琪,苏建家,黄定瑞,等.人乙型肝炎病毒感染与原发性肝癌:在树鼩的实验研究.肝脏病杂志, 1994, 2(4): 196-199.
51覃柳亮,苏建家,杨春,等.树鼩实验性肝癌过程中GGT酶阳性肝细胞灶、IGF-Ⅱ及HBxAg异常表达.广西医学, 1998, 20(1): 4-7.
52苏建家,严瑞琪,覃国忠,等.树鼩实验肝癌发生过程中IGF-Ⅱ、HBxAg、p21和p53蛋白的表达.临床与实验病理学杂志, 1998, 14(5): 484-487.
53班克臣,苏建家,杨春,等.在HBV和(或)AFB1诱发树树鼩肝癌过程中肝组织ras癌基因的表达.实验动物科学与管理, 1999, 16(1): 4-7.
54李瑗,苏建家,严瑞琪,等.乙肝病毒和黄曲霉毒素B1在树树鼩肝癌形成中的协同作用.癌症, 1999, 18(2): 129-132.
55李瑗,苏建家,严瑞琪,等. HBV和AFB1引起树树鼩肝癌形成过程中IGF-Ⅱ蛋白表达的动态观察.广西医科大学学报, 1999, 16(4): 395-398.
56苏建家,李瑗,班克臣,等.动态研究HBV和AFB1诱发树鼩肝细胞癌过程中一些基因的表达.广西医科大学学报, 2001, 18(2): 151-154.
57李瑗,苏建家,曹骥,等.用cDNA阵列技术研究黄曲霉毒素B1诱发树鼩肝癌形成过程中的基因变化.中华肝脏病杂志, 2003, 11(2): 96-99.
58苏建家,李瑗,班克臣,等.树树鼩实验性肝癌发生过程p53基因的变化.中华肝脏病杂志, 2003, 11(3): 159-161.
59李瑗,苏建家,曹骥,等.不同因素诱发的树树鼩肝癌组织的基因表达差异.癌症, 2003, 22(10): 1018-1022.
60段小娴,欧盛敬,李瑗,等.树鼩肝癌形成过程中凋亡抑制基因的表达及与细胞凋亡的关系.中国临床康复, 2004, 8(2): 392-393.
61 Park US, Su JJ, Ban KC,et al. Mutations in the p53 tumor suppressor gene in tree shrew hepatocellular carcinoma associated with hepatitis B virus infection and intake of aflatoxin B1. Gene, 2000, 251(1):73-80.
62苏建家.实验性肝细胞癌研究状况.医学文选, 2001, 20(1): 1-3.
63李瑗, Karen Baumgartner,Denise MacMillan,等.树鼩的人工哺育.上海实验动物科学, 2001, 21(2): 70-74.
1 Wasinger VC,Cordwell SJ,Cerpa-Poljak A,et al.Progress with gene-product mapping of the Mollicutes:Mycoplasma genitalium. Electrophoresis, 1995, 16(7): 1090-1094.
2马吉湘,曾常茜.在肝细胞癌诊断中的肿瘤标志物.北华大学学报(自然科学版), 2003, 4(5): 382-385.
3 Sipmson RJ,Dorow DS.Cancer proteomics:from signaling networks to tumor markers.Trends Biotechnol, 2001, 19(10 suppl): 40-48.
4 Chevalier S,Macdonald N,Tonge R,et al. Proteomic analysis of differential protein expression in primary hepatocytes induced by EGF,tumor necrosis factor alpha or the peroxisome proliferator nafenopin.Eur J Biiochem, 2000, 267(15):4624-4634.
5 Jungblut PR, Zimny-Arndt U, Zeindl-Eberhart E, et al. Proteomics in humandisease: cancer, heart and infectious diseases. Electrophoresis, 1999, 20(10): 2100-2110.
6俞利荣,王楠,吴高德,等.人肝癌细胞系BEL-7404和正常肝细胞系L-02表达的蛋白质组双向凝胶电泳分析.科学通报, 2000, 45(2): 170-178.
7 Yu LR, Zeng R, Shao XX,et al. Identification of differentially expressed proteins between human hepatoma and normal liver cell lines by two-dimensional electrophoresis and liquid chromatography-ion trap mass spectrometry. Electrophoresis, 2000, 21(14): 3058-3068.
8曾嵘,俞利荣,邵晓霞,等.电喷雾离子阱质谱法鉴定人肝癌细胞双向凝胶电泳胶内蛋白质.科学通报, 2000, 45(6): 592-598.
9 Park KS, Cho SY, Kim H, et al. Proteomic alterations of the variants of human aldehyde dehydrogenase isozymes correlate with hepatocellular carcinoma. Int J Cancer, 2002, 97(2): 261-265.
10 Park KS, Kim H, Kim NG, et al. Proteomic analysis and molecular characterization of tissue ferritin light chain in hepatocellular carcinoma. Hepatology, 2002, 35(6): 1459-1466.
11 Yu LR, Shao XX, Jiang WL, et al. Proteome alterations in human hepatoma cells transfected with antisense epidermal growth factor receptor sequence. Electrophoresis, 2001, 22(14): 3001-3008.
12刘博,黄志强,周宁新,等.肝癌细胞系SMMC7721蛋白质组学初步分析.中华实验外科杂志, 2004, 21(12): 1439-1440.
13 Yokoyama Y, Kuramitsu Y, Takashima M, et al. Proteomic profiling of proteins decreased in hepatocellular carcinoma from patients infected with hepatitis C virus. Proteomics, 2004, 4(7): 2111-2116.
14 Li C, Hong Y, Tan YX,et al. Accurate qualitative and quantitative proteomic analysis of clinical hepatocellular carcinoma using laser capture microdissection coupled with isotope-coded affinity tag and two-dimensional liquid chromatography mass spectrometry. Mol Cell Proteomics, 2004, 3(4): 399-409.
15 Fujii K, Kondo T, Yokoo H, et al. Proteomic study of human hepatocellular carcinoma using two-dimensional difference gel electrophoresis with saturation cysteine dye. Proteomics, 2005, 5(5): 1411-1422.
16彭小宁,范学工,黄燕,等.螺杆菌感染与肝癌关系的研究.世界华人消化杂志, 2002, 10(8): 902-906.
17黄燕,范学工,陈永平,等.肝癌组织中螺杆菌16SrRNA基因的检测.世界华人消化杂志,2002,10(8):877-882.
18 Dore MP,Realdi G,Mura D,et al.Helicobacter infection in patients with HCV-related chronic hepatitis,cirrhosis,and hepatocellular carcinoma. Dig Dis Sci, 2002, 47(7): 1638-1643.
19 Pellicano R,Mazzaferro V,Grigioni WF,et al. Helicobacter species sequences in liver samples from patients with and without hepatocellular carcinoma. World J Gastroenterol, 2004, 10(4): 598-601.
20 Ito K,Nakamura M,Toda G,et al.Potential role of helicobacter pylori in hepatocarcinogenesis. Int J Mol med, 2004, 13(2): 221-227.
21张艳,范学工,陈仁,等.幽门螺杆菌对人肝细胞系HepG2细胞蛋白质组的影响.世界华人消化杂志, 2004, 12(6): 1325-1328.
22 Yuan Q, An J, Liu DG, et al. Proteomic analysis of differential protein expression in a human hepatoma revertant cell line by using an improved two-dimensional electrophoresis procedure combined with matrix assisted laser desorption/ionization-time of flight-mass spectrometry. Electrophoresis, 2004, 25(7-8): 1160-1168.
23 Seow TK, Ong SE, Liang RC,et al. Two-dimensional electrophoresis map of the human hepatocellular carcinoma cell line, HCC-M, and identification of the separated proteins by mass spectrometry. Electrophoresis, 2000, 21(9): 1787-1813.
24 Seow TK, Liang RC, Leow CK, et al. Hepatocellular carcinoma: from bedside to proteomics. Proteomics, 2001, 1(10): 1249-1263.
25 Ou K, Seow TK, Liang RC,et al. Proteome analysis of a human heptocellularcarcinoma cell line, HCC-M: an update. Electrophoresis, 2001, 22(13): 2804- 2811.
26 Liang RC, Neo JC, Lo SL,et al. Proteome database of hepatocellular carcinoma. J Chromatogr B Analyt Technol Biomed Life Sci, 2002, 771(1-2): 303-328.
27胡志远,万晶宏,王利红,等.蛋白质组研究中双向聚丙烯酰胺凝胶电泳图谱的计算机分析.军事医学科学院院刊, 1998, 22(4): 301-304.
28 Chew EC, Liew CT, Wu S, et al. Electrophoretic analysis of nuclear matrix proteins in human hepatocellular carcinoma. Anticancer Res, 1997, 17(5A): 3581-3585.
29 Yokoo H, Kondo T, Fujii K, et al. Proteomic signature corresponding to alpha fetoprotein expression in liver cancer cells. Hepatology, 2004, 40(3): 609-617.
30肖雪媛,刘丹慧,刘博,等.体外培养的不同亚型肝癌细胞差异蛋白的初步筛查.解剖学报, 2004, 35(6): 602-606.
31 Bottoni P, Giardina B, Martorana GE, et al. A two dimensional electrophoresis preliminary approach to human hepatocarcinoma differentiation induced by PPAR-agonists. J Cell Mol Med, 2005, 9(2): 462- 467.
32 Srisomsap C, Sawangareetrakul P, Subhasitanont P, et al. Proteomic analysis of cholangiocarcinoma cell line. Proteomics, 2004, 4(4): 1135-44.
33 Anderson NL, Anderson NG. The human plasma proteome: history, character, and diagnostic prospects.Mol Cell Proteomics, 2002, 1(11): 845- 867.
34 Kennedy S.Proteomic profiling from human samples: the body fluid alternative. Toxicol Lett, 2001,120(1-3): 379-384.
35 Poon TC, Johnson PJ. Proteome analysis and its impact on the discovery of serological tumor markers. Clin Chim Acta, 2001, 313(1-2): 231-239.
36 Poon TC, Yip TT, Chan AT,et al. Comprehensive proteomic profilingidentifies serum proteomic signatures for detection of hepatocellular carcinoma and its subtypes. Clin Chem, 2003, 49(5): 752-760.
37 Steel LF, Shumpert D, Trotter M,et al. A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma. Proteomics, 2003, 3(5): 601-609.
38王征,阮幼冰,官阳.肝细胞癌患者血清蛋白质组成分的双向凝胶电泳-飞行时间质谱分析.中华病理学杂志, 2003, 32(4): 333-336.
39宋森涛,朱梁,王文静,等.应用SELDI-TOF MS技术分析肝癌血清差异表达蛋白.实用癌症杂志, 2004, 19(6): 601-604.
40 Shalhoub P, Kern S, Girard S, et al. Proteomic-based approach for the identification of tumor markers associated with hepatocellular carcinoma. Dis Markers, 2001, 17(4): 217-223.
41 Le Naour F, Brichory F, Misek DE,et al. A distinct repertoire of autoantibodies in hepatocellular carcinoma identified by proteomic analysis. Mol Cell Proteomics. 2002,1(3):197-203.
42 Comunale MA,Mattu TS, Lowman MA, et al. Comparative proteomic analysis of de-N-glycosylated serum from hepatitis B carriers reveals polypeptides that correlate with disease status.Proteomics, 2004, 4(3): 826- 838.
43 Schwegler EE, Cazares L, Steel LF, et al. SELDI-TOF MS profiling of serum for detection of the progression of chronic hepatitis C to hepatocellular carcinoma. Hepatology,2005,41(3):634-642.
44 Paradis V, Degos F, Dargere D, et al. Identification of a new marker of hepatocellular carcinoma by serum protein profiling of patients with chronic liver diseases. Hepatology, 2005, 41(1): 40-47.
45 Yang ZF, Ho DW, Lam CT, et al. Identification of brain-derived neurotrophic factor as a novel functional protein in hepatocellular carcinoma. Cancer Res, 2005, 65(1): 219-225.
46 Block TM, Comunale MA, Lowman M, et al. Use of targetedglycoproteomics to identify serum glycoproteins that correlate with liver cancer in woodchucks and humans. Proc Natl ACad Sci U S A, 2005, 102(3): 779-784.
47 Ueda K.Proteome analysis of autoantibodies in sera of patients with cancer.Rinsho Byori, 2005, 53(5):437-445.
48 Hippo Y, Watanabe K, Watanabe A, et al. Identificantion of soluble NH2-terminal fragment of glypican-3 as a serological marker for early-stage hepatocellular carcinoma. Cancer Res, 2004, 64(7):2418-2423.
49 Zeindl-Eberhart E, Klugbauer S, Dimitrijevic N, et al. Proteome analysis of rat hepatomas: carcinogen-dependent tumor-associated protein variants. Electrophoresis, 2001, 22(14):3009-3018.
50 Zeindl-Eberhart E, Haraida S, Liebmann S, et al. Detection and identification of tumor-associated protein variants in human hepatocellular carcinomas. Hepatology, 2004, 39(2): 540-549.
51 Lim SO, Park SJ, Kim W, et al. Proteome analysis of hepatocellular carcinoma. Biochem Biophys Res Commun, 2002, 291(4): 1031-1037.
52 Kim J, Kim SH, Lee SU, et al. Proteome analysis of human liver tumor tissue by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of disease-related proteins.Electrophoresis, 2002, 23(24): 4142-4156.
53 Kim W, Oe Lim S, Kim JS, et al. Comparison of proteome between hepatitis B virus- and hepatitis C virus-associated hepatocellular carcinoma. Clin Cancer Res, 2003, 9(15): 5493-5500.
54 Tannapfel A, Anhalt K, Hausermann P, et al. Identification of novel proteins associated with hepatocellular carcinomas using protein microarrays. J Pathol, 2003, 201(2): 238-249.
55 Melle C, Kaufmann R, Hommann M, et al. Proteomic profiling in micro -dissected hepatocellular carcinoma tissue using ProteinChip technology. Int J Oncol, 2004,24(4):885-891.
56龙云铸,范学工,李宁,等.应用双向电泳对HBV相关性肝细胞癌肝组织蛋白质组的初步研究.中国医师杂志, 2003, 5(6): 731-732.
57 Takashima M, Kuramitsu Y, Yokoyama Y, et al. Proteomic profiling of heat shock protein 70 family members as biomarkers for hepatitis C virus-related hepatocellular carcinoma. Proteomics, 2003, 3(12): 2487-2493.
58吴珊,刘宗石,邱殷庆,等.人原发性肝细胞癌核基质蛋白的研究.中华肿瘤杂志, 1997, 19(5): 339-341.
59 Liang CR, Leow CK, Neo JC, et al. Proteome analysis of human hepatocellular carcinoma tissues by two-dimensional difference gel electrophoresis and mass spectrometry. Proteomics, 2005, 5(8): 2258-2271.
60 Li C, Tan YX, Zhou H, et al. Proteomic analysis of hepatitis B virus-associated hepatocellular carcinoma:Identification of potential tumor markers. Proteomics, 2005, 5(4): 1125-1139.
61 Takashima M, Kuramitsu Y, Yokoyama Y, et al. Overexpression of alpha enolase in hepatitis C virus-related hepatocellular carcinoma: association with tumor progression as determined by proteomic analysis. Proteomics, 2005, 5(6): 1686-1692.
62 Le Naour F.Contribution of proteomics to tumor immunology. Proteomics, 2001, 1(10): 1295-1302.
63 Ding SJ, Li Y, Tan YX, et al. From proteomic analysis to clinical significance: overexpression of cytokeratin 19 correlates with hepatocellular carcinoma metastasis. Mol Cell Proteomics, 2004, 3(1): 73-81.
64 Nagai T,Murota M, Nishioka M, et al. Elevation of cytokeratin 19 fragment in serum in patients with hepatoma: its clinical significance. Eur J Gastroenterol Hepatol, 2001, 13(2): 157-161.
65 Ding SJ, Li Y, Shao XX, et al. Proteome analysis of hepatocellular carcinoma cell strains, MHCC97-H and MHCC97-L, with different metastasis potentials. Proteomics, 2004, 4(4): 982-994.
66崔杰峰,刘银坤,宋海燕,等.不同转移潜能人肝癌细胞系的差异表达蛋白质组分析及意义.中华检验医学杂志, 2004, 27(4): 240-244.
67宋海燕,刘银坤,崔杰峰,等.人肝细胞性肝癌组织转移相关分子的比较蛋白质组学研究.中华肝脏病杂志, 2005, 13(5): 331-334.
68张丽君,崔杰峰,刘银坤,等.不同转移潜能人肝癌细胞系酪氨酸磷酸化蛋白质差异分析.中华肝脏病杂志, 2005, 13(6): 436-4393.
69叶丽虹,秦宵然,张晓东,等.原肌球蛋白、波形纤维蛋白和热休克蛋白70在肝癌转移亚细胞中表达上调.中国生物化学与分子生物学报, 2005, 21(2): 244-249.
70代智,刘银坤,崔杰峰,等.肝癌转移相关的核心岩藻糖基化蛋白质表达谱的研究.生物化学与生物物理进展, 2005, 32(5):442-448.
71 Page MJ, Amess B, Rohlff C, et al. Proteomics: a major new technology for the drug discovery process. Drug Discov Today, 1999, 4(2): 55-62.
72 Ong ES, Len SM, Lee AC. Differential protein expression of the inhibitory effects of a standardized extract from Scutellariae radix in liver cancer cell lines using liquid chromatography and tandem mass spectrometry. J Agric Food Chem, 2005, 53(1): 8-16.
73 Luk JM, Su YC, Lam SC, et al. Proteomic identification of Ku70/Ku80 autoantigen recognized by monoclonal antibody against hepatocellular carcinoma. Proteomics, 2005, 5(7): 1980-1986.
74 Steel LF, Mattu TS, Mehta A, et al. A proteomic approach for the discovery of early detection markers of hepatocellular carcinoma. Dis Markers, 2001, 17(3): 179-189.
75龙云铸,范学工,李宁,等.应用肝组织蛋白质组模式诊断乙型肝炎病毒相关性肝细胞癌的初步研究.中华肝脏病杂志, 2004, 12(4): 231-233.
76黄成,樊嘉,周俭,等.肝细胞癌门静脉癌栓形成的血清蛋白质分子标记物研究.中华医学杂志, 2005, 85(11): 781-785.
77罗治文.蛋白质芯片质谱技术在肿瘤蛋白质组学中应用的研究进展.实用癌症杂志, 2004, 19(4): 429-431, 439.
78贺福初.国家人类肝脏蛋白质组计划.医学研究通讯, 2004, 33(7): 2.
2. Tang ZY, Ye SL, Liu YK, et al. A decade’s studies on metastasis of hepatocellular carcinoma. J Cancer Res Clin Oncol, 2004, 130(4): 187-196.
3.张思维,李连弟,鲁凤珠,等.中国1990-1992年原发性肝癌死亡调查分析.中华肿瘤杂志, 1999, 21(4): 245-249.
4. Summers J, Smolec JM, Snyder R. A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci USA, 1978, 75(9): 4533-4537.
5. Marion PL, Van Davelaar MJ, Knight SS, et al. Hepatocellurlar carcinoma in ground squirrels persistently infected with ground squirrel hepatitis virus. Proc Natl Acad Sci USA, 1986, 83(12): 4543-4546.
6. Cora L, Wild CP, Mehrotra R, et al. Contribution of aflatoxin B1 and hepatitis B virus infection in the induction of liver tumors in ducks. Cancer Res, 1990, 50(7): 2156- 2163.
7. Kim CM, Koike K, Saito J, et al. HBx gene of hepatitis B virus induces liver cancer in transgenic mice. Nature, 1992, 315(6324): 317-320.
8. Prince AM, Brotman B. Perspectives of hepatitis B studies with chimpanzees. ILAR J, 2001, 42: 85-88.
9.苏建家,严瑞琪,甘友全,等.成年树鼩实验感染人乙型肝炎病毒的研究.中华病理学杂志, 1987, 16: 103-106.
10.王荷英,刘季和,喻林冲,等.树鼩实验麻风的组织病理学观察.中华皮肤科杂志, 1996, 29(4): 258-260.
11.万新邦,庞其方,丘福禧,等.成年的中国云南树鼩对人轮状病毒易感性的实验研究.中华微生物学和免疫学杂志, 1985, 5(5): 304-307.
12.曾怀德,柴慧霞,谢扬高,等.马桑内酯引起的树鼩实验性癫痫.四川医学院学报, 1984, (2): 159-163.
13.梁真,冼苏,林健,等.树鼩糖尿病模型肝脏功能及形态学变化与TNF-α的关系.广西医科大学学报, 2003, 20(5): 639-641.
14.苏建家,李瑗,班克臣,等.树树鼩实验性肝癌发生过程p53基因的变化.中华肝脏病杂志, 2003, 11(3): 159-161.
15. Yan RQ, Su JJ,Huang DR, et al. Human hepatitis B Virus and hepatocellular carcinoma I Experimental infection of tree shrews with hepatitis B Virus. J Cancer Res clin onco, 1996, 122(5): 283-288.
16. Yan RQ, Su JJ, Huang DR, Gan YC, yang C. Huang GH. Human hepatitis B Virus and hepatocellular cainoma. II Experimental induction of hepatocllular carcinoma in tree shrews exposed to hepatitis B virus and aflatoxin B1. J Cancer Clin Oncol, 1996, 122(5): 289-295.
17.李瑗,苏建家,覃柳亮,等.乙肝病毒和黄曲霉毒素B1诱发树鼩肝癌的病理变化.临床与实验病理学杂志, 2000, 16(3): 224-227.
18. Li Y, Su JJ, Qi LL,et al. Synergistic effect of hepatitis B Virus and aflatoxin B1 in hepatocarcinogenesis in tree shrews. Ann Acad Med Singapore, 1999, 28(1): 67-71.
19.李瑗,苏建家,曹骥,等.不同因素诱发的树鼩肝癌组织的基因表达差异.癌症, 2003, 22(10): 1018-1022.
20.何琼琼,程瑞雪,陈主初.蛋白质组学在肝癌研究中的应用.国外医学肿瘤学分册, 2005, 32(10): 782-784.
21.许克新,王云川.双向电泳及质谱分析与蛋白质组研究.第四军医大学学报, 2002, 23(22): 2017-2022.
22. O’Farrel PH. High resolution Two-dimensional electrophoresis of proteins. J Bilo chem, 1975, 20:4007.
1.郭鹞.人类疾病的动物模型.第一版.北京:人民卫生出版社, 1990: 1-6.
2.彭燕章,叶智彰,邹如金,等.树鼩生物学.昆明:云南科技出版社, 1991: 1-4.
3.严瑞琪,苏建家,陈志英,等.人乙型肝炎病毒实验感染成年树鼩的初步研究.广西医学院学报, 1984, 1(1): 10-15.
4.苏建家.实验性肝细胞癌研究状况.医学文选, 2001, 20(1): 1-3.
5. Brewer NR. Personalities in the early history of laboratory animal science and medicine.LabAnisci, 1980, 30(4): 741-758.
6. Crafton TS. The founding and early history of the national society of medical research. Lab Anisci, 1980, 30(40): 760-764.
7. Park US, SuJJ,Ban KC, et al. Mutations in the p53 tumoe suppressor gene in tree shrew hepatocellular carcinoma associate with hepatitis Bvirus infection and intake of aflatoxin B1.Gene, 2000, 251(1): 73-80.
8.李瑗,苏建家,曹骥,等.用cDNA阵列技术研究黄曲霉素B1诱发树鼩肝癌过程中的基因变化.中华肝脏病杂志, 2003, 11(2): 96-99.
9.詹美云,刘崇柏,李成明,等.甲型肝炎病毒感染树鼩的初步研究.中国医学科学院学报, 1981, 3(3): 148-152.
10.王凤水,陈红松,王宇,等.树鼩感染人乙肝病毒动物模型研究中的非特异性反应.北京医科大学学报, 1996, 28(5): 340-342.
11.刘志,毛青,王宇明,等.丙型肝炎病毒感染成年树鼩的实验研究.第三军医大学学报, 1998, 20(6): 472-475.
12.李奇芬,丁明权,王洪,等.成年树鼩实验感染丁型肝炎病毒的研究.第三军医大学学报, 1994, 16(4): 234.
13.李奇芬,毛青,吴纯清,等.树鼩实验性丁型肝炎的病理观察.中华肝脏病杂志, 1996, 4(4): 210-213.
14.李奇芬,毛青,吴纯清,等.树鼩传代感染HDV后肝脏内抗原表达及病理变化.世界华人消化杂志, 1999, 7(5): 378-381.
15.万新邦,庞其方,丘福禧,等.成年的中国云南树鼩对人轮状病毒易感性的实验研究.中华微生物学和免疫学杂志, 1985, 5(5): 304-307.
16. Darai G, Zoller L, Matz B, et al. Experimental infection and the state of viral latency of adult Tupaia with herpes simplex virus type 1 and 2 and infection of juvenile Tupaia with temperature-sensitive mutants of HSV type 2. Arch Virol, 1980, 65(3-4): 311-318.
17.戴长柏.人单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染树鼩的研究.医学生物学研究, 1984, 1: 62-69.
18.朱宇同,王维祥,曾子安,等.流感病毒感染成年树鼩的研究.上海实验动物科学, 1986, 6(6): 20-23.
19.刘丽华,李正平,俞永新.树鼩对登革热病毒敏感性的研究.中华微生物学和免疫学杂志, 1985, 5(5): 329-330.
20.侯宗柳,白登云,黄文丽,等.树鼩对森林脑炎病毒的敏感性及发病机理的研究.中国病毒学, 1991, 6(4): 298-303.
21.鲍铭晖,钱小榆,张海林,等.树鼩感染基孔肯雅病毒的病理学研究.云南医药, 1992, 13 (3): 180-182.
22.张海林,米竹青,施华芳,等.树鼩实验感染后血清中抗基孔肯雅病毒IgM和IgG抗体检查.地方病通报, 1997, 12(2): 26-27.
23. McBrien NA, Cornell LM, Gentle A. Structural and ultrastructural changes to the sclera in a mammalian model of high myopia. Invest Ophthalmol Vis Sci, 2001, 42(10): 2179-2187.
24. Gentle A, Liu Y, Martin JE, Conti GL, et al. Collagen gene expression and the altered accumulation of scleral collagen during the development of high myopia. J Biol Chem, 2003, 278(19): 16587-16594.
25. Fuchs E, Flugge G.Psychosocial stress induces molecular and structural alterations in the brain - How animal experiments help to understand pathomechanisms of depressive illnesses. Psychosom Med Psychother, 2001, 47(1): 80-97.
26. Fuchs E. Social stress in tree shrews as an animal model of depression: an example of a behavioral model of a CNS disorder. CNS Spectr, 2005, 10(3): 182-189.
27.苏建家,李瑗,班克臣,等.树鼩实验性肝癌发生过程p53基因的变化.中华肝脏病杂志, 2003, 11(3): 159-161.
28. Ishiko S, Yoshida A, Mori F,et al. Early ocular changes in a tree shrew model of diabetes.Nippon Ganka Gakkai Zasshi, 1997, 101(1): 19-23.
29. Steinhausen M, Thederan H, Nolinski D,et al. Acute renal failure in Tupaia belangeri and rats. Bibl Anat, 1979, (18): 90-92.
30.曾怀德,柴慧霞,谢扬高,等.马桑内酯引起的树鼩实验性癫痫.四川医学院学报, 1984, 2: 159-163.
31. Schwaier A. Tupaias (tree shrews)--a new animal model for gallstone research; first observations of gallstones. Res Exp Med (Berl), 1979, 176(1):15-24.
32.谈跃,王子灿,李麟仙.树鼩局部脑缺血模型的研究.昆明医学院学报, 1994, 15(3): 17-21.
33. Braun H, Kloft W. Hamatologische untersuchngen am spizhornchcn Tupata lana.Tupaiidae Prosimiae, 1965, 21: 663-664.
34. Hunt RD, Chalifoux L. The hemogram of the tree shrew. Folia Priimat, 1967, 7: 34-36.
35.邹如金,代伟,昆龙,等.树鼩血相的研究.动物学研究, 1983, 4(3): 291-293.
36.李瑗, Karen Baumgartner, Denise MacMillan,等.树鼩的人工哺育.上海实验动物科学, 2001, 21(2): 70-74.
1.许克新,王云川.双向电泳及质谱分析与蛋白质组研究.第四军医大学学报, 2002, 23(22): 2017-2022.
2.郭尧君.SDS电泳技术的实验考虑及最新进展.生物化学与生物物理学进展, 1991, 18: 32-35.
3. O’Farrel PH. High resolution Two-dimensional electrophoresis of proteins.J Bilo chem, 1975, 20: 4007.
4. Cordwell SJ, Nouwens AS, Verrills NM,et al. Subproteomics based upon protein cellular location and relative solubilities in conjunction with composite two-dimensional dldtrophoredid gels. Electrophoresis, 2000, 21:1094-1103.
5. Gorg A, Obermaier C, Boguth G,et al. The current state of two-dimensional eletrophoresis with immobilized pH gradients.Eletrophoresis, 2000, 32: 1037 -1053.
6.朱雪琼,石一夏,陈怀增,等.子宫肌瘤蛋白质组研究中二维电泳方法的建立.浙江大学学报(医学版), 2004, 6(33): 550-552.
7.郭尧君.载体两性电解质pH梯度等电聚焦.见:郭尧君,主编.蛋白质电泳实验技术.第一版.北京:科学出版社出版, 1999, 161-210.
8. Fountoulakis M, Takacs B. Effect of strong detergents and chaotropes on the Deiection of proyeins in two-dimensional gels.Eletrophoresis, 2001, 22(9): 1593-1602.
9. Molloy MP. Two-dimensional eletrophoresis of membrane proteins using immobilizsd pH gradients.Anal Biochem, 2000, 280(1): 1-10.
10. Rabillooud T, Adessi C, Giraudel A, et al. Improcement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients. Eletrophoresis, 1997, 18: 307-316.
11. Herbert BR, Molloy MP, Gooley AA, et al. Improved protein solubility in two-dimensional electrophoresis using tributyl phosphine as reducing agent.Electrophoresis, 1998, 19: 845-851.
12. Gorg A,Obermaier C,Boguth G,et al.The current state of two-dimensional electrophoresis with immobilized pH gradients.Electrophoresis, 2000, 21(6): 1037-1053.
13. Qian xiao-hong.Proteomics: from Protein Sequence to Function. Beijing: Scientific Publisher, 2002, 57-58.
14. Yan JX, Wait R, Berkelman T, et al. A moditidf silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ioni-zation and electrospray ionization-mass spectrometry. Electrophresis, 2000, 21(17): 3666-3667.
15.Brush M. Dye haed: protein gel staining products.Scientific, 1998, 12: 16-22.
16.Neuhoff V, Arold N, Taube D, Ehrhardt W. Improved staining of proteins in polyacrylamide gels including isorlrctric focusing Gels with clear background at nanogram sensitivity using Coomassir Brilliant Blue G-250 and R-250.Electrophoresis, 1988, 9(6): 255-262.
17.Neuhoff V, Stamm R, Pardowitz I, et al. Essential problems in quantification og preteins following cloolidal staining with coomassie brilliant blue dyes in polyacrylamide gels, and rheir solution.Electrophoresis, 1990, 11(2): 101- 117.
1.王树声,苏建家,张华远,等.树鼩、熊猴感染人乙型肝炎病毒血清指症的研究.中华实验和临床病毒学杂志, 2003, 17(1): 21-24.
2. Kim J, Kim SH, Lee SU, et al. Proteome analysis of human liver tumor tissue by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of disease-related proteins. Electrophoresis, 2002, 23(24): 4142-4156.
3. Cha C, Dematteo RP. Molecular mechanisms in hepatocellular carcinoma development. Best Pract Res Clin Gastroenterol, 2005, 19(1): 25-37
4. Ming L, Thorgeirsson SS, Gail MH, et a1. Dominant role of hepatitis B virusand cofactor role of aflatoxin in hepatocarcinogenesis in qidong China. Hepatotology, 2002, 36(5): 1214-1220
5. Groopman JD, Wang JS, Scholl P. Molecular biomarkers for aflatoxins:from adducts to gene mutations to human liver cancer.Can J Physiol Pharmacol, 1996, 4(2): 203-209.
6.苏建家,李瑗,班克臣,等.动态研究HBV和AFB1诱发树鼩肝细胞癌过程中一些基因的表达,广西医科大学报, 2001, 18(2): 151-154.
7. Garrido C, Galluzzi L, Brunet M, et al. Mechanisms of cytochrome c release from mitochondria.Cell Death and Differentiation , 2006,5: 1–11.
8. Yanagawa T, Omura K, Harada H, et a1.Peroxiredoxin I expression in tongue squamous cell carcinomas as involved in tumor recurrence. Int J Oral Maxillofac Surg, 2005, 34(8): 915-920.
9. Shen J, Person MD, Zhu J,et al. Protein expression profiles in pancreatitis as detected by two-dimensional gel electrophoresis and mass spectrometry. Cancer Res, 2004, 64(24): 9018-9026.
10. Kim HJ, Chae HZ, Kim YJ, et a1.Preferential elevation of Prx I and Trx expression in lung cancer cells following hypoxia and in human lung cancer tissues.Cell Biol Toxicol, 2003, 19(5): 285-298.
11. Crowley-Weber CL, Payne CM, Gleason-Guzman M, et a1.Development and molecular characterization of HCT-116 cell lines resistant to the tumor promoter and multiple stress-inducer deoxycholate. Carcinogenesis,2002, 23(12): 2063-2080.
12. Choi JH,Kim TN,Kim S,et a1.Overexpression of mitochondrial thioredoxin reductase and peroxiredoxin III in hepatocellular carcinomas. Anticancer Res, 2002, 22(6A): 3331-3335.
13. Jang JS, Cho HY, Lee YJ, et al. The differential proteome profile of stomach cancer: identification of the biomarker candidates.Oncol Res, 2004, 14(10): 491-499.
14. Yong-Zheng Wu, Yefim Manevich, James L, et al. Interaction of SurfactantProtein A with Peroxiredoxin 6 Regulates Phospholipase A2 Activity.Biol. Chem, 2006, 11(281): 7515-7525.
15. Miyazawa1 T, Kubo1 E, Fatma N, et al. Peroxiredoxin6–Deficient Mouse Lens Epithelial Cells Exhibit an Increased Susceptibility to Hypergylycemia Induced Apoptosis. Invest Ophthalmol Vis Sci, 2005(46): 3859.
16. Xuesong Ouyang, Theodore L, DeWeese, et al. Loss-of-Function of Nkx3.1 Promotes Increased Oxidative Damage in Prostate Carcinogenesis. Cancer Research, 2005(65): 6773-6779.
17.周海君,刘银坤,崔杰峰,等.人肝癌细胞系的糖蛋白质组学研究.生物化学与生物物理进展, 2006, 33(1): 59-64.
18. Varesco L, Caligo MA, Simi P, et al. The NM23 gene maps to human chromosome band 17q22 and shows a restriction fragment length polymorphism with BgⅠ,Ⅱ.Genes Chromosom Cancer, 1992, 4(1):84.
19. Urano T, Furuhawa K, Sbihu H. Expression of NM23/NDP kinase proteins on the cell surface.Oncogene, 1993, 8: 1371.
20. Leone A, Flatow U, King CR, et al. Reduced tumor incidence, metastatic potential and cytokine responsiveness of NM23-transfected melanoma cell.Cell, 1991, 65(1): 25-35.
21. Ennessy C, Henry JA, May FEB, et al. Expression of the antimetastatic gene NM23 in human breast cancer. J Natl Cancer Inst, 1991, 83: 281.
22. Irayama R, Sawai S, Takagi Y, et al. Positive relationship between expression of anti-metastatic factor (NM23 gene product or nucleotide diphosphate kinase) and good prognosis in human breast cancer. J NatI Cancer Inst, 1991, 83: 1249.
23. Nakayama H,Yasui W,Yokozaki H,et al. Reduced expression of NM23 is associated with metastasis of human gastric carcinomas. Jpn J Cancer Res, 1993, 84(2): 184-190.
24. Müller W, Schneiders A, Hommel G, et al. Expression of NM23 in gastric carcinoma:association with tumor progression and poor prognosis1. Cancer,1998, 83(12): 2481.
25. Yamaguchi A, Urano T, Fushida S, et al. Inverse association of NM23-H1 expression by colorectal cancer with liver metastasis.Br J Cancer 1993, 68(5): 1020-1024.
26. Nakayama T, Ohtsuru A,Nakao K, et al. Expression in human hepatocellular carcinoma of nucleotide diphosphate kinase,a homologue of the NM23 gene product. J Natl Cancer Inst, 1992, 84:1349.
27. Rosengard AM, Krutzsch HC, Sbearn A, et al. Reduced NM23/and protein in tumor metastasis and abberant drosphila development.Nature, 1989, 342: 177.
28.盛红光,张敏,姜言明,等.肿瘤血管形成和转移抑制基因NM 23与肝细胞癌侵袭性关系的研究.中华普通外科, 2004, 19(3): 180.
29. Lombardi D, Lacombe ML, Paggi MG. NM23:unraveling its biological function in cell differentiation.J Cell Physio, 2000, 82(2): 144.
30. Caligo MA, Cipollini G, Fiore L, et al. NM23 gene expression correlates with cell growth rate and sphate. Int J Cancer, 1995, 60(6): 837.
31. Kantor JD, McCormick B, Steeg PS, et al. Inhibition of cell mortality after NM23 transfection of human and murine tumor cell.Cancer Res, 1993, 53 (9): 1971.
32.张豪,孙桂菊.乙肝病毒和化学致癌物致肝细胞癌变的研究进展.肿瘤, 2004, 24(5): 509-511.
33.李瑗,苏建家,曹骥,等.用cDNA阵列技术研究黄曲霉毒素B1诱发树鼩肝癌形成过程中的基因变化.中华肝脏病杂志, 2003, 11(2): 96-99.
34. Allred DC, Avis I, et a1.Diferentialexpressionofthe early lung can cer detection marker, heterogeneous nuclear ribonucleopmtein-A2/B1 innormal breast abreastand neoplastic breastcancer.Breast CancerRes,2001, 66: 217-224.
35. Wu S, Sato M, Endo C, Sakurada A, et a1. hnRNP B1 protein may be a possible prognostic factor in squamous cell carcinoma of the lung. LungCancer, 2003, 41(2): 179-186.
36. Satoh H, Kamma H, Ishikawa H, et a1.Expression of hnRNP A2/B1 proteins in human cancer cell lines. Int J Oncol, 2000, 16(4): 763-767.
37. Ushigome M, Ubagai T, Fukuda H, et a1.Up-regulation of hnRNP A1 gene in sporadic human colorectal cancers. Int J Oncol, 2005, 26(3): 635-640.
38. Christian K, Lang M, et a1. Interaction of heterogeneous nuclear ribonucleoprotein A1 with cytochrome P450 CYP2A6 mRNA: implications for post-transcriptional regulation of the CYP2A6 gene. Mol Pharmacol, 2004, 65(6): 1405-1414.
39. Kamma H, FujimotoM, FujiwaraM, et a1. Interactionofhn-RNP A2/B1 isoforms with telomefic ssDNA and the invitro function.Biochem Biophy Res Comm, 2001, 280: 625-630.
40. Iwanaga K, Sueoka N, Sato A,et al.Heterogeneous nuclear ribonucleoprotein B1 protein impairs DNA repair mediated through the inhibition of DNA-dependent protein kinase activity. Biochem Biophys Res Commun, 2005, 333(3): 888-895.
41. Jang JS, Cho HY, Lee YJ, et al. The differential proteome profile of stomach cancer: identification of the biomarker candidates. Oncol Res, 2004, 14 (10): 491-499.
42. Gehrmann ML,Fenselau C,Hathout Y. Highly altered protein expression profile in the adriamycin resistant MCF-7 cell line. J Proteome Res, 2004, 3(3): 403-409.
43. Jimin Xu, Marco Capezzone, Xiao Xu, et al. Activation of Nicotinamide N-Methyltransferase Gene Promoter by Hepatocyte Nuclear Factor-1? in Human Papillary Thyroid Cancer Cells. Molecular Endocrinology, 2005, 19(2): 527-539
44. Narimatsu T,Tamori A,Koh N,et a1.p16 promoter hypermethylation in human hepatocellular carcinoma with or without hepatitis virus infection. Intervirology, 2004, 47(1): 26-31.
45.吴曼,薛开先,孙开来,等.肿瘤遗传学.北京:科学出版社.2004: 3-26.
46. Pang R, Yuen J, Yuen MF, et a1. PIN 1 overexpression and beta-catenin gene mutations are distinct oneogenie events in human hepatocellular carcinoma. Oneogene, 2004, 23 (23): 4182-4186.
47. Zhong S,Yeo W,Tang MW, et a1.Intensive hyperm ethylation of the CpG island of Ras association domain family lA in hepatitis B virus-associated hepatocellular carcinomas.Clin Cancer Res, 2003, 9(9): 3376-3382.
48. Heilbronn R, Sehlehofer JR, Yalkinoglu AO, et al. Selective DNA- amplification induced by carcinogens (initiators): evidence for a role of proteases and DNA polymerase alpha. Int J Cancer, 1985, 36(1): 85-91.
49. Zhang YJ, Ahsan H, Chen Y, et a1. High frequency of promoter hyperm ethylation of RASSF1A and p16 and its relationship to aflatoxin B1-DNA adduct levels in human hepatocellular carcinoma. Mol Carcinog, 2002, 35(2): 85-92.
50. Sohn S,Jaitovitch-Groisman I,Benlimame N,et a1.Retroviral expression of the hepatitis B virus X gene promotes liver cell susceptibility to carcinogen-induced site specific mutagenesis.Mutat Res,2000,460(1):17.
51. Livezey KW, Negorev D, Simon D, et a1.Increased chromosomal alterations and micronuclei formation in human hepatoma HepG2 cells transfected with the hepatitis B virus HBX gene. Mutat Res, 2002, 505(1-2): 63.
52. Capovilla A, Arbuthnot P. Hepatitis B virus X protein does not influence essential steps of nucleotide excision repair effected by human liver extracts. Biochem Biophys Res Commun, 2003, 312(3): 806.
53. Wild CP, Shrestha SM, Anwar WA, er al. Field studies of aflatoxin exposure, metabolism and induction of genetic alterations in relation to HBV infection and hepatocellular carcinoma in The Gambia and Thailand. Toxicol Lett, 1992, 64-65: 455-461.
54. Bontron S, Lin-Marq, Strubin M, et a1. Hepatitis B virus X proteinassociated with UV-DDB1 induces cell death in the nucleus and is functionally antagonized by UV-DDB2.J Biol Chem,2002,277(41):3884
55. liams CS,Dubois RN. Prostaglandin endoperoxide synthase: Why two isoforms .Am J Physiol, 1996, 270(1): 393.
56. Sadrzadeh SM, Nanji AA. The 212aminosteroid 162desmethyl tirilazad mesylate prevents necroinflammatory changes in experimental alcoholic liver disease. J Pharmacol Exp Ther, 1998, 284(1): 406.
57. Nanji AA, Miao L, Thomas P, et al. Enhanced cyclooxygenase gene expression in alcoholic liver disease in the rat. Gastroenterology, 1997, 112 (3): 943.
58. Nieto N, Greenwel P, Friedman SL, et al. Ethanol and arachidonic acid increase alpha 2(I)collagen expression in rat hepatic stellate cells overexpressing cytochrome P450 2El; role of H2O2 and cyclooxygenase-2.J Biol Chem, 2000, 275(26): 20136-20145.
59. Weiss P, Mouallem M, Bruck R, et al. Nimesulide2induced hepatitis and acute liver failure.Isr Med Assoc J , 1999, 1(2): 89.
60. Shiota G, Okubo M, Noumi T, et al. Cyclooxygenase expression in hepatocel lularcarcionoma.Hepatogastroenterology, 1999, 46(25): 407.
61. Koga H, Sakisaka S, Ohishi M, et al. Expression of COX-2 in human hepatocellular carcinoma : relevance to tumor dedifferentation . Hepatology, 1999, 29(3): 688
62. Choi JH, Kim TN, Kim S, et a1. Overexpression of mitochondrial thioredoxin reductase and peroxiredoxinⅢin hepatocellular carcinomas. Anticancer Res, 2002, 22(6A): 3331-3335.
63. Haapasalo H, Kylaniemi M, Paunul N, et a1. Expression of antioxi-dant enzymes in astrocytic brain tumors. Brain Pathol, 2003, 13(2): 155-164.
64. Lincoln DT, Ali Emadi EM, Tonissen KF, et a1.The thioredoxin -thioredoxin reductase system: over-expression in human cancer. Anticancer Res, 2003, 23(3B): 2425-2433.
65. Gromer S,Gross JH.Methylseleninate is a substrate rather than an inhibitor of mammalian thioredoxin reductase.Implications for the antitumor effects of selenium. J Biol Chem, 2002, 277(12): 9701-9706.
66. Edamoto Y,Hara A,Biernat W,et a1. Alterations of RB1, p53 and W nt pathways in hepatocellular carcinomas associated with hepatitis C,hepatitis B and alcohol liver cirrhosis. Int J Cancer, 2003, 106(3): 334-341.
67. Szabo E,Paska C,Kaposi Novak P,et a1.Similarities and difierences in hepatitis B an d C virus induced hepatocarcinogenesis.Pathol Oncol Res,2004, 10(1): 5-11. .
1. Rhee SG, Kang SW, Chang TS, et al. peroxiredoxin, a novel family of peroxi -dases. IUBMB Life, 2001, 52(1-2): 35-41.
2. Fujii J, Ikeda Y. Advances in our understanding of peroxiredoxin, a multi- functional,mammalian redox protein. Redox Rep, 2002, 7(3): 123-130.
3. Tien Nguyen-nhu N, Knoops B. Mitochondrial and cytosolic expression of human peroxiredoxin 5 in Saccharomyces cerevisiae protect yeast cells from oxidative stress induced by paraquat. FEBS Lett, 2003, 544: 148-152.
4. ChangT S,Jeong W,Choi SY,et al.Regulation of peroxiredoxin I activity by Cdc2-mediated phosphorylation. J Biol Chem, 2002, 277(28):25370- 25376.
5. Wood Z A, Schroder E, Robin Harris J, et a1. Structure, mechanisnl and regulation of peroxiredoxins.Trends Biochem Sc1, 2003, 28 (1): 32-40
6. Fujii J, Ikeda Y. Advances in our understanding of peroxiredoxin, a multi -functional, mammalian redox protein. Redox Rep, 2002, 7(3):123-130.
7. Henkle-Duhrsen K, Kanapkotter A.Antioxidant enzvyme families in parasitic nematodes. Mol Biochem Parasitol, 200l, 114(2): l29-l42.
8. Rabilloud T, Heller M, Gasnier F, et a1. Proteonfics analvsis of cellular response to oxidative stress.Evidence for in vivo overoxidation of peroxire -doxins at their active site. J Biol Chem, 2002, 277(22): l9396-1940l.
9. Zhou Y,Kok K H,Chun A C,et al.Mouse peroxiredoxin V is a:thioredoxin peroxidase that inhibits p53-induced apoptosis.Biochem Biophys Res Commun, 2000, 268(3): 921-927.
10. Shan H,Butterfield L H,Chiu R,et al.Cloning and sequeuce analysis of candidate human natural killer-enhancing factor genes. Immunogenetics, 1994, 40(2): 129-134.
11.章波,向渝梅,白云.抗氧化蛋白Peroxiredoxin家族研究进展.生理科学进展, 2004, 35(4): 352-355.
12.周海君,刘银坤,崔杰峰,等.人肝癌细胞系的糖蛋白质组学研究.生物化学与生物物理进展, 2006, 33(1): 59-64.
13. Crowley-Weber CL, Payne CM, Gleason-Guzman M, et a1.Development and molecular characterization of HCT-116 cell lines resistant to the tumor promoter an d multiple stress-inducer,deoxycholate.Carcinogenesis, 2002, 23: 2063-2080.
14. Kim HJ, Chae HZ, Kim YJ, et a1. Preferential elevation of Prx I and Trx expression in lung cancer cells following hypoxia and in human lung cancer tissues.Cell Biol Toxicol, 2003, 19(5): 285-298.
15. Jang JS, Cho HY, Lee YJ, et al. The differential proteome profile of stomach cancer: identification of the biomarker candidates.Oncol Res, 2004, 14(10): 491-499.
16. Chang TS, Jeong W, Choi SY,et al. Regulation ofPeroxiredoxin I Activity by Cdc2-mediated Phosphorylation .J Biol Chem, 2002, 277(28): 25370-25376.
17. Gisin J, Perren A, Bawohl M, Rare allelic imbalances, but no mutations of the PRDX1 gene in human hepatocellular carcinomas. Journal of Clinical Pathology, 2005, 58(11): 1229- 1231.
18. Choi MH, Lee IK, Kim GW, et al. Regulation of PDGF signalling and vascular remodelling by peroxiredoxin II. Nature, 2005, 435(7040): 347-353.
19. Kim H, Lee TH, Park ES, et al. Role of peroxiredoxins in regulating intra -cellular hydrofen peroxide and hydrogen peroxide-induced apoptosis in thyroid cell. J Biol Chen, 2000, 275(24): 18266-18270.
20.王世鄂,黄威权,付建芳等. Peroxiredoxin II在小鼠卵巢中卵泡、GV卵、MII卵和植入前胚的表达和定位.解剖学报, 2004, 35(6): 647-650.
21. Araki M.Nanri H.Ejima K,et al.Antioxidant function of the mitochondrial protein SP-22 in the cardiovascular systen1. J Biol Chem, l999, 274(4): 227l -2278.
22. Shih S F,Wu Y H,Hung CH,et a1.Abrin triggers cell death by inactivating a thiol-specific antioxidant protein. J Biol Chem, 200l, 276 (24): 2l870-2l877.
23. Masaki M,Ikeda A,Shiraki E,et al.Mixed lineage kinase LZK and anti -oxidant protein-l activate NF-kappaB synergistically.Eur J Biochem, 2003, 270(1): 76-83.
24. Jasehke A.Mi H.Fropschug M.Human T cell cyelophilin 18 binds to thiol- specific antioxidant protein Aopl and stimulates its activity. J Mol Biol, 1998, 277(4): 763-769.
25.冯艳,刘东青,王震,等.心肌特异性新蛋白激酶p93和peroxiredoxin 3的相互作用.生物化学与生物物理进展, 2004, 31(8): 688-691.
26. Larisa Nonn1, Margareta Berggren, et al. Increased Expression of Mitochondrial Peroxiredoxin-3 (Thioredoxin Peroxidase-2) Protects Cancer Cells Against Hypoxia and Drug-Induced Hydrogen Peroxide-Dependent Apoptosis1. Molecular Cancer Research, 2003, 1: 682-689.
27. Tong-Shin Chang, Chun-Seok Cho, Sunjoo Park, et al. Peroxiredoxin III, a Mitochondrion-specific Peroxidase, Regulates Apoptotic Signaling byMitochondria. J Biol Chem, 2004, 279(40): 41975-41984.
28. Kamma H,FujimotoM,FujiwaraM,et a1.Interactionofhn-RNP A2/B1 isoforms with telomefic ssDNA and the invitro function.Biochem Biophy Res Comm, 2001, 280:625-630.
29. Miyazawa1 T, Kubo1 E,Fatmay N,et al.Peroxiredoxin6–Deficient Mouse Lens Epithelial Cells Exhibit an Increased Susceptibility to Hypergylycemia Induced Apoptosis. Invest Ophthalmol Vis Sci, 2005(46): 3859.
30. Yan Wang, Shelley A. Phelan, Yefim Manevich, Sheldon I. Transgenic Mice Overexpressing Peroxiredoxin 6 Show Increased Resistance to Lung Injury in Hyperoxia. American Journal of Respiratory Cell and Molecular Biology, 2006(34): 481-486.
1詹美云,刘崇柏,李成明,等.甲型肝炎病毒感染树鼩的初步研究.中国医学科学院学报, 1981, 3 (3): 148-152.
2严瑞琪,苏建家,陈志英,等.人乙型肝炎病毒实验感染成年树鼩的初步研究.广西医学院学报, 1984, 1(1): 10-15.
3苏建家,严瑞琪,甘友全,等.成年树鼩实验感染人乙型肝炎病毒的研究.中华病理学杂志, 1987, 16(2): 103-106.
4王凤水,陈红松,王宇,等.树鼩感染人乙肝病毒动物模型研究中的非特异性反应.北京医科大学学报, 1996, 28(5): 340-342.
5王海平,姚志强,周永兴,等.树鼩实验感染乙型肝炎病毒的再评价.中华肝脏病杂志, 1998, 6(2): 96-97.
6苏建家,李瑗,王树声,等.树鼩对人乙型肝炎病毒易感性的验证.广西医学, 2000, 7(4): 298-301.
7王树声,苏建家,张华远,等.树鼩、熊猴感染人乙型肝炎病毒血清指征的研究.中华实验和临床病毒学杂志, 2003, 17(1): 21-24.
8王海平,周永兴,姚志强,等.成年树鼩实验感染丙型肝炎病毒的初步研究.第四军医大学学报, 1997, 18(4): 375-376.
9刘志,毛青,王宇明,等.丙型肝炎病毒感染成年树鼩的实验研究.第三军医大学学报,1998,20(6): 472-475.
10 Xie ZC, Riezu-Boj JI, Lasarte JJ, et al. Transmission of hepatits C virus infection to tree shrews. Virology, 1998, 244(2): 513-520.
11谢志春,高伟志,苏洁寒,等.树鼩对丙型肝炎病毒的易感性研究.广西医科大学学报, 2000, 17(3): 347-350.
12 Zhao X, Tang ZY, Klumpp B,et al. Primary hepatocytes of Tupaia belangeri as a potential model for hepatitis C virus infection. J Clin Invest. 2002,109(2):221-32.
13李奇芬,丁明权,王洪,等.成年树鼩实验感染丁型肝炎病毒的研究.第三军医大学学报, 1994, 16(4): 234.
14李奇芬,毛青,吴纯清,等.树鼩实验性丁型肝炎的病理观察.中华肝脏病杂志, 1996, 4(4): 210-213.
15李奇芬.毛青.吴纯清.等.树鼩传代感染HDV后肝脏内抗原表达及病理变化.世界华人消化杂志, 1999, 7(5): 378-381.
16谢志春,周元平,吴继周,等. TTV感染成年树鼩的初步研究.广西预防医学, 2000, 6(3): 133-135.
17万新邦,刘玑昌,胥爱源,等.用人轮状病毒感染成年树鼩的实验研究.中华医学杂志, 1982, 62 (8): 461-465.
18万新邦.中国云南成年树鼩对人轮状病毒的易感性的实验研究.医学生物学研究, 1985, 1: 1-10
19万新邦,庞其方,丘福禧,等.成年的中国云南树鼩对人轮状病毒易感性的实验研究.中华微生物学和免疫学杂志, 1985, 5(5): 304-307.
20 Darai G, Zoller L, Matz B,et al.Experimental infection and the state of viral latency of adult Tupaia with herpes simplex virus type 1 and 2 and infection of juvenile Tupaia with temperature-sensitive mutants of HSV type 2.Arch Virol, 1980, 65(3-4): 311-318.
21徐维明.人单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染树鼩的超微结构研究.医学生物学研究, 1984, (1): 73-77.
22戴长柏.人单纯疱疹病毒Ⅱ型(HSV-Ⅱ)感染树鼩的研究.医学生物学研究, 1984,(1): 62-69.
23朱宇同,王维祥,曾子安,等.流感病毒感染成年树鼩的研究.上海实验动物科学, 1986, 6(6): 20-23.
24刘丽华,李正平,俞永新.树鼩对登革热病毒敏感性的研究.中华微生物学和免疫学杂志, 1985, 5(5): 329-330.
25侯宗柳,白登云,黄文丽,等.树鼩对森林脑炎病毒的敏感性及发病机理的研究.中国病毒学, 1991, 6(4): 298-303.
26鲍铭晖,钱小榆,张海林,等.树鼩感染基孔肯雅病毒的病理学研究.云南医药, 1992, 3: 180-182.
27张海林,米竹青,施华芳,等.树鼩实验感染后血清中抗基孔肯雅病毒IgM和IgG抗体检查.地方病通报, 1997, 12(2): 26-27.
28王荷英,叶顺章,刘季和,等.麻风菌接种树鼩的研究.中国麻风杂志, 1989, 5(3): 137-139.
29薛文昌,韩永高,陈剑英.等.麻风杆菌已经能在树鼩足垫中生长:麻风杆菌又一个新的动物模型.皮肤病与性病, 1990, 12(1): 1-5.
30王荷英,刘季和,喻林冲,等.树鼩实验麻风的组织病理学观察.中华皮肤科杂志, 1996, 29(4): 258-260.
31王志光,朱杰玲,武云,等.树鼩人工感染马锥虫的研究.上海实验动物科学, 1987, 7(2):79.
32 Ishiko S, Yoshida A, Mori F,et al. Early ocular changes in a tree shrew model of diabetes. Nippon Ganka Gakkai Zasshi, 1997, 101(1): 19-23.
33彭葆坤,周宁,唐哲,等.实验性糖尿病树鼩四周心肌超微结构改变和心脏血液动力学变化.中华临床医药杂志, 2004, 5(4): 8-11.
34梁真,冼苏,林健,等.树鼩糖尿病模型肝脏功能及形态学变化与TNF-α的关系.广西医科大学学报, 2003, 20(5): 639-641.
35谈跃,王子灿,李麟仙.树鼩局部脑缺血模型的研究.昆明医学院学报, 1994, 15(3): 17-21.
36王子灿,李麟仙,谈跃,等.树鼩局部脑缺血模型病理生理特点及实验治疗的研究.中国病理生理杂志, 1997,13(5): 492.
37 Steinhausen M, Thederan H, Nolinski D,et al. Acute renal failure in Tupaia belangeri and rats. Bibl Anat. 1979,(18):90-2.
38曾怀德,柴慧霞,谢扬高,等.马桑内酯引起的树鼩实验性癫痫.四川医学院学报, 1984, 2 : 159-163.
39 Schwaier A. Tupaias (tree shrews)--a new animal model for gallstone research; first observations of gallstones. Res Exp Med (Berl), 1979, 176(1):15-24.
40佘铬鹏,夏人仪,王宗立,等.树鼩高密度脂蛋白(HDL)抗动脉粥样硬化的研究:一种新的动物模型.医学研究通讯, 1985, 3 : 89-90.
41 Zadnik K,Mutti DO.How applicable are animal myopia models to human juvenile onset myopia?Vision Res, 1995, 35(9): 1283-1288.
42 Edwards MH.Animal models of myopia.A review.Acta Ophthalmol Scand, 1996, 74(3): 213-219.
43 McBrien NA, Cornell LM, Gentle A. Structural and ultrastructural changes to the sclera in a mammalian model of high myopia.. Invest Ophthalmol Vis Sci, 2001, 42(10): 2179-2187.
44 Gentle A, Liu Y, Martin JE, Conti GL,et al. Collagen gene expression and the altered accumulation of scleral collagen during the development of high myopia. J Biol Chem, 2003, 278(19): 16587-16594.
45 Fuchs E, Flugge G. Psychosocial stress induces molecular and structural alterations in the brain - How animal experiments help to understand pathomechanisms of depressive illnesses Psychosom Med Psychother. 2001, 47(1): 80-97.
46 Czeh B, Michaelis T, Watanabe T,et al. Stress-induced changes in cerebral metabolites, hippocampal volume, and cell proliferation are prevented by antidepressant treatment with tianeptine. Proc Natl ACad Sci U S A, 2001 , 98(22): 12796-12801.
47 Fuchs E, Flugge G. Social stress in tree shrews: effects on physiology, brain function, and behavior of subordinate individuals. Pharmacol Biochem Behav, 2002 ,73(1):247-258.
48 Fuchs E. Social stress in tree shrews as an animal model of depression: an example of a behavioral model of a CNS disorder. CNS Spectr. 2005 , 10(3): 182-189.
49严瑞琪,苏建家,黄定瑞,等.人乙型肝炎病毒和黄曲霉毒素B1诱发树鼩原发性肝癌的研究.中华病理学杂志, 1989, 18(1): 19-22.
50严瑞琪,苏建家,黄定瑞,等.人乙型肝炎病毒感染与原发性肝癌:在树鼩的实验研究.肝脏病杂志, 1994, 2(4): 196-199.
51覃柳亮,苏建家,杨春,等.树鼩实验性肝癌过程中GGT酶阳性肝细胞灶、IGF-Ⅱ及HBxAg异常表达.广西医学, 1998, 20(1): 4-7.
52苏建家,严瑞琪,覃国忠,等.树鼩实验肝癌发生过程中IGF-Ⅱ、HBxAg、p21和p53蛋白的表达.临床与实验病理学杂志, 1998, 14(5): 484-487.
53班克臣,苏建家,杨春,等.在HBV和(或)AFB1诱发树树鼩肝癌过程中肝组织ras癌基因的表达.实验动物科学与管理, 1999, 16(1): 4-7.
54李瑗,苏建家,严瑞琪,等.乙肝病毒和黄曲霉毒素B1在树树鼩肝癌形成中的协同作用.癌症, 1999, 18(2): 129-132.
55李瑗,苏建家,严瑞琪,等. HBV和AFB1引起树树鼩肝癌形成过程中IGF-Ⅱ蛋白表达的动态观察.广西医科大学学报, 1999, 16(4): 395-398.
56苏建家,李瑗,班克臣,等.动态研究HBV和AFB1诱发树鼩肝细胞癌过程中一些基因的表达.广西医科大学学报, 2001, 18(2): 151-154.
57李瑗,苏建家,曹骥,等.用cDNA阵列技术研究黄曲霉毒素B1诱发树鼩肝癌形成过程中的基因变化.中华肝脏病杂志, 2003, 11(2): 96-99.
58苏建家,李瑗,班克臣,等.树树鼩实验性肝癌发生过程p53基因的变化.中华肝脏病杂志, 2003, 11(3): 159-161.
59李瑗,苏建家,曹骥,等.不同因素诱发的树树鼩肝癌组织的基因表达差异.癌症, 2003, 22(10): 1018-1022.
60段小娴,欧盛敬,李瑗,等.树鼩肝癌形成过程中凋亡抑制基因的表达及与细胞凋亡的关系.中国临床康复, 2004, 8(2): 392-393.
61 Park US, Su JJ, Ban KC,et al. Mutations in the p53 tumor suppressor gene in tree shrew hepatocellular carcinoma associated with hepatitis B virus infection and intake of aflatoxin B1. Gene, 2000, 251(1):73-80.
62苏建家.实验性肝细胞癌研究状况.医学文选, 2001, 20(1): 1-3.
63李瑗, Karen Baumgartner,Denise MacMillan,等.树鼩的人工哺育.上海实验动物科学, 2001, 21(2): 70-74.
1 Wasinger VC,Cordwell SJ,Cerpa-Poljak A,et al.Progress with gene-product mapping of the Mollicutes:Mycoplasma genitalium. Electrophoresis, 1995, 16(7): 1090-1094.
2马吉湘,曾常茜.在肝细胞癌诊断中的肿瘤标志物.北华大学学报(自然科学版), 2003, 4(5): 382-385.
3 Sipmson RJ,Dorow DS.Cancer proteomics:from signaling networks to tumor markers.Trends Biotechnol, 2001, 19(10 suppl): 40-48.
4 Chevalier S,Macdonald N,Tonge R,et al. Proteomic analysis of differential protein expression in primary hepatocytes induced by EGF,tumor necrosis factor alpha or the peroxisome proliferator nafenopin.Eur J Biiochem, 2000, 267(15):4624-4634.
5 Jungblut PR, Zimny-Arndt U, Zeindl-Eberhart E, et al. Proteomics in humandisease: cancer, heart and infectious diseases. Electrophoresis, 1999, 20(10): 2100-2110.
6俞利荣,王楠,吴高德,等.人肝癌细胞系BEL-7404和正常肝细胞系L-02表达的蛋白质组双向凝胶电泳分析.科学通报, 2000, 45(2): 170-178.
7 Yu LR, Zeng R, Shao XX,et al. Identification of differentially expressed proteins between human hepatoma and normal liver cell lines by two-dimensional electrophoresis and liquid chromatography-ion trap mass spectrometry. Electrophoresis, 2000, 21(14): 3058-3068.
8曾嵘,俞利荣,邵晓霞,等.电喷雾离子阱质谱法鉴定人肝癌细胞双向凝胶电泳胶内蛋白质.科学通报, 2000, 45(6): 592-598.
9 Park KS, Cho SY, Kim H, et al. Proteomic alterations of the variants of human aldehyde dehydrogenase isozymes correlate with hepatocellular carcinoma. Int J Cancer, 2002, 97(2): 261-265.
10 Park KS, Kim H, Kim NG, et al. Proteomic analysis and molecular characterization of tissue ferritin light chain in hepatocellular carcinoma. Hepatology, 2002, 35(6): 1459-1466.
11 Yu LR, Shao XX, Jiang WL, et al. Proteome alterations in human hepatoma cells transfected with antisense epidermal growth factor receptor sequence. Electrophoresis, 2001, 22(14): 3001-3008.
12刘博,黄志强,周宁新,等.肝癌细胞系SMMC7721蛋白质组学初步分析.中华实验外科杂志, 2004, 21(12): 1439-1440.
13 Yokoyama Y, Kuramitsu Y, Takashima M, et al. Proteomic profiling of proteins decreased in hepatocellular carcinoma from patients infected with hepatitis C virus. Proteomics, 2004, 4(7): 2111-2116.
14 Li C, Hong Y, Tan YX,et al. Accurate qualitative and quantitative proteomic analysis of clinical hepatocellular carcinoma using laser capture microdissection coupled with isotope-coded affinity tag and two-dimensional liquid chromatography mass spectrometry. Mol Cell Proteomics, 2004, 3(4): 399-409.
15 Fujii K, Kondo T, Yokoo H, et al. Proteomic study of human hepatocellular carcinoma using two-dimensional difference gel electrophoresis with saturation cysteine dye. Proteomics, 2005, 5(5): 1411-1422.
16彭小宁,范学工,黄燕,等.螺杆菌感染与肝癌关系的研究.世界华人消化杂志, 2002, 10(8): 902-906.
17黄燕,范学工,陈永平,等.肝癌组织中螺杆菌16SrRNA基因的检测.世界华人消化杂志,2002,10(8):877-882.
18 Dore MP,Realdi G,Mura D,et al.Helicobacter infection in patients with HCV-related chronic hepatitis,cirrhosis,and hepatocellular carcinoma. Dig Dis Sci, 2002, 47(7): 1638-1643.
19 Pellicano R,Mazzaferro V,Grigioni WF,et al. Helicobacter species sequences in liver samples from patients with and without hepatocellular carcinoma. World J Gastroenterol, 2004, 10(4): 598-601.
20 Ito K,Nakamura M,Toda G,et al.Potential role of helicobacter pylori in hepatocarcinogenesis. Int J Mol med, 2004, 13(2): 221-227.
21张艳,范学工,陈仁,等.幽门螺杆菌对人肝细胞系HepG2细胞蛋白质组的影响.世界华人消化杂志, 2004, 12(6): 1325-1328.
22 Yuan Q, An J, Liu DG, et al. Proteomic analysis of differential protein expression in a human hepatoma revertant cell line by using an improved two-dimensional electrophoresis procedure combined with matrix assisted laser desorption/ionization-time of flight-mass spectrometry. Electrophoresis, 2004, 25(7-8): 1160-1168.
23 Seow TK, Ong SE, Liang RC,et al. Two-dimensional electrophoresis map of the human hepatocellular carcinoma cell line, HCC-M, and identification of the separated proteins by mass spectrometry. Electrophoresis, 2000, 21(9): 1787-1813.
24 Seow TK, Liang RC, Leow CK, et al. Hepatocellular carcinoma: from bedside to proteomics. Proteomics, 2001, 1(10): 1249-1263.
25 Ou K, Seow TK, Liang RC,et al. Proteome analysis of a human heptocellularcarcinoma cell line, HCC-M: an update. Electrophoresis, 2001, 22(13): 2804- 2811.
26 Liang RC, Neo JC, Lo SL,et al. Proteome database of hepatocellular carcinoma. J Chromatogr B Analyt Technol Biomed Life Sci, 2002, 771(1-2): 303-328.
27胡志远,万晶宏,王利红,等.蛋白质组研究中双向聚丙烯酰胺凝胶电泳图谱的计算机分析.军事医学科学院院刊, 1998, 22(4): 301-304.
28 Chew EC, Liew CT, Wu S, et al. Electrophoretic analysis of nuclear matrix proteins in human hepatocellular carcinoma. Anticancer Res, 1997, 17(5A): 3581-3585.
29 Yokoo H, Kondo T, Fujii K, et al. Proteomic signature corresponding to alpha fetoprotein expression in liver cancer cells. Hepatology, 2004, 40(3): 609-617.
30肖雪媛,刘丹慧,刘博,等.体外培养的不同亚型肝癌细胞差异蛋白的初步筛查.解剖学报, 2004, 35(6): 602-606.
31 Bottoni P, Giardina B, Martorana GE, et al. A two dimensional electrophoresis preliminary approach to human hepatocarcinoma differentiation induced by PPAR-agonists. J Cell Mol Med, 2005, 9(2): 462- 467.
32 Srisomsap C, Sawangareetrakul P, Subhasitanont P, et al. Proteomic analysis of cholangiocarcinoma cell line. Proteomics, 2004, 4(4): 1135-44.
33 Anderson NL, Anderson NG. The human plasma proteome: history, character, and diagnostic prospects.Mol Cell Proteomics, 2002, 1(11): 845- 867.
34 Kennedy S.Proteomic profiling from human samples: the body fluid alternative. Toxicol Lett, 2001,120(1-3): 379-384.
35 Poon TC, Johnson PJ. Proteome analysis and its impact on the discovery of serological tumor markers. Clin Chim Acta, 2001, 313(1-2): 231-239.
36 Poon TC, Yip TT, Chan AT,et al. Comprehensive proteomic profilingidentifies serum proteomic signatures for detection of hepatocellular carcinoma and its subtypes. Clin Chem, 2003, 49(5): 752-760.
37 Steel LF, Shumpert D, Trotter M,et al. A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma. Proteomics, 2003, 3(5): 601-609.
38王征,阮幼冰,官阳.肝细胞癌患者血清蛋白质组成分的双向凝胶电泳-飞行时间质谱分析.中华病理学杂志, 2003, 32(4): 333-336.
39宋森涛,朱梁,王文静,等.应用SELDI-TOF MS技术分析肝癌血清差异表达蛋白.实用癌症杂志, 2004, 19(6): 601-604.
40 Shalhoub P, Kern S, Girard S, et al. Proteomic-based approach for the identification of tumor markers associated with hepatocellular carcinoma. Dis Markers, 2001, 17(4): 217-223.
41 Le Naour F, Brichory F, Misek DE,et al. A distinct repertoire of autoantibodies in hepatocellular carcinoma identified by proteomic analysis. Mol Cell Proteomics. 2002,1(3):197-203.
42 Comunale MA,Mattu TS, Lowman MA, et al. Comparative proteomic analysis of de-N-glycosylated serum from hepatitis B carriers reveals polypeptides that correlate with disease status.Proteomics, 2004, 4(3): 826- 838.
43 Schwegler EE, Cazares L, Steel LF, et al. SELDI-TOF MS profiling of serum for detection of the progression of chronic hepatitis C to hepatocellular carcinoma. Hepatology,2005,41(3):634-642.
44 Paradis V, Degos F, Dargere D, et al. Identification of a new marker of hepatocellular carcinoma by serum protein profiling of patients with chronic liver diseases. Hepatology, 2005, 41(1): 40-47.
45 Yang ZF, Ho DW, Lam CT, et al. Identification of brain-derived neurotrophic factor as a novel functional protein in hepatocellular carcinoma. Cancer Res, 2005, 65(1): 219-225.
46 Block TM, Comunale MA, Lowman M, et al. Use of targetedglycoproteomics to identify serum glycoproteins that correlate with liver cancer in woodchucks and humans. Proc Natl ACad Sci U S A, 2005, 102(3): 779-784.
47 Ueda K.Proteome analysis of autoantibodies in sera of patients with cancer.Rinsho Byori, 2005, 53(5):437-445.
48 Hippo Y, Watanabe K, Watanabe A, et al. Identificantion of soluble NH2-terminal fragment of glypican-3 as a serological marker for early-stage hepatocellular carcinoma. Cancer Res, 2004, 64(7):2418-2423.
49 Zeindl-Eberhart E, Klugbauer S, Dimitrijevic N, et al. Proteome analysis of rat hepatomas: carcinogen-dependent tumor-associated protein variants. Electrophoresis, 2001, 22(14):3009-3018.
50 Zeindl-Eberhart E, Haraida S, Liebmann S, et al. Detection and identification of tumor-associated protein variants in human hepatocellular carcinomas. Hepatology, 2004, 39(2): 540-549.
51 Lim SO, Park SJ, Kim W, et al. Proteome analysis of hepatocellular carcinoma. Biochem Biophys Res Commun, 2002, 291(4): 1031-1037.
52 Kim J, Kim SH, Lee SU, et al. Proteome analysis of human liver tumor tissue by two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-mass spectrometry for identification of disease-related proteins.Electrophoresis, 2002, 23(24): 4142-4156.
53 Kim W, Oe Lim S, Kim JS, et al. Comparison of proteome between hepatitis B virus- and hepatitis C virus-associated hepatocellular carcinoma. Clin Cancer Res, 2003, 9(15): 5493-5500.
54 Tannapfel A, Anhalt K, Hausermann P, et al. Identification of novel proteins associated with hepatocellular carcinomas using protein microarrays. J Pathol, 2003, 201(2): 238-249.
55 Melle C, Kaufmann R, Hommann M, et al. Proteomic profiling in micro -dissected hepatocellular carcinoma tissue using ProteinChip technology. Int J Oncol, 2004,24(4):885-891.
56龙云铸,范学工,李宁,等.应用双向电泳对HBV相关性肝细胞癌肝组织蛋白质组的初步研究.中国医师杂志, 2003, 5(6): 731-732.
57 Takashima M, Kuramitsu Y, Yokoyama Y, et al. Proteomic profiling of heat shock protein 70 family members as biomarkers for hepatitis C virus-related hepatocellular carcinoma. Proteomics, 2003, 3(12): 2487-2493.
58吴珊,刘宗石,邱殷庆,等.人原发性肝细胞癌核基质蛋白的研究.中华肿瘤杂志, 1997, 19(5): 339-341.
59 Liang CR, Leow CK, Neo JC, et al. Proteome analysis of human hepatocellular carcinoma tissues by two-dimensional difference gel electrophoresis and mass spectrometry. Proteomics, 2005, 5(8): 2258-2271.
60 Li C, Tan YX, Zhou H, et al. Proteomic analysis of hepatitis B virus-associated hepatocellular carcinoma:Identification of potential tumor markers. Proteomics, 2005, 5(4): 1125-1139.
61 Takashima M, Kuramitsu Y, Yokoyama Y, et al. Overexpression of alpha enolase in hepatitis C virus-related hepatocellular carcinoma: association with tumor progression as determined by proteomic analysis. Proteomics, 2005, 5(6): 1686-1692.
62 Le Naour F.Contribution of proteomics to tumor immunology. Proteomics, 2001, 1(10): 1295-1302.
63 Ding SJ, Li Y, Tan YX, et al. From proteomic analysis to clinical significance: overexpression of cytokeratin 19 correlates with hepatocellular carcinoma metastasis. Mol Cell Proteomics, 2004, 3(1): 73-81.
64 Nagai T,Murota M, Nishioka M, et al. Elevation of cytokeratin 19 fragment in serum in patients with hepatoma: its clinical significance. Eur J Gastroenterol Hepatol, 2001, 13(2): 157-161.
65 Ding SJ, Li Y, Shao XX, et al. Proteome analysis of hepatocellular carcinoma cell strains, MHCC97-H and MHCC97-L, with different metastasis potentials. Proteomics, 2004, 4(4): 982-994.
66崔杰峰,刘银坤,宋海燕,等.不同转移潜能人肝癌细胞系的差异表达蛋白质组分析及意义.中华检验医学杂志, 2004, 27(4): 240-244.
67宋海燕,刘银坤,崔杰峰,等.人肝细胞性肝癌组织转移相关分子的比较蛋白质组学研究.中华肝脏病杂志, 2005, 13(5): 331-334.
68张丽君,崔杰峰,刘银坤,等.不同转移潜能人肝癌细胞系酪氨酸磷酸化蛋白质差异分析.中华肝脏病杂志, 2005, 13(6): 436-4393.
69叶丽虹,秦宵然,张晓东,等.原肌球蛋白、波形纤维蛋白和热休克蛋白70在肝癌转移亚细胞中表达上调.中国生物化学与分子生物学报, 2005, 21(2): 244-249.
70代智,刘银坤,崔杰峰,等.肝癌转移相关的核心岩藻糖基化蛋白质表达谱的研究.生物化学与生物物理进展, 2005, 32(5):442-448.
71 Page MJ, Amess B, Rohlff C, et al. Proteomics: a major new technology for the drug discovery process. Drug Discov Today, 1999, 4(2): 55-62.
72 Ong ES, Len SM, Lee AC. Differential protein expression of the inhibitory effects of a standardized extract from Scutellariae radix in liver cancer cell lines using liquid chromatography and tandem mass spectrometry. J Agric Food Chem, 2005, 53(1): 8-16.
73 Luk JM, Su YC, Lam SC, et al. Proteomic identification of Ku70/Ku80 autoantigen recognized by monoclonal antibody against hepatocellular carcinoma. Proteomics, 2005, 5(7): 1980-1986.
74 Steel LF, Mattu TS, Mehta A, et al. A proteomic approach for the discovery of early detection markers of hepatocellular carcinoma. Dis Markers, 2001, 17(3): 179-189.
75龙云铸,范学工,李宁,等.应用肝组织蛋白质组模式诊断乙型肝炎病毒相关性肝细胞癌的初步研究.中华肝脏病杂志, 2004, 12(4): 231-233.
76黄成,樊嘉,周俭,等.肝细胞癌门静脉癌栓形成的血清蛋白质分子标记物研究.中华医学杂志, 2005, 85(11): 781-785.
77罗治文.蛋白质芯片质谱技术在肿瘤蛋白质组学中应用的研究进展.实用癌症杂志, 2004, 19(4): 429-431, 439.
78贺福初.国家人类肝脏蛋白质组计划.医学研究通讯, 2004, 33(7): 2.