摘要
目的:注射法建立金黄地鼠颊囊癌模型,动态观测肿瘤发生进程;探讨碳离子束辐照对机体微量元素以及凋亡相关基因Survivin、Caspase-3表达的影响,为重离子肿瘤治疗和空间辐射危险评估提供实验依据。
材料与方法:1.本研究选取81只健康的金黄地鼠随机分为3组,其中空白对照组(6只),涂抹组(15只),注射组(60只)(又分A, B, C, D, E5个小组)。采用预设浓度为(0.5%,0.4%,0.3%,0.2%,0.1%)的DMBA丙酮注射法和涂抹法(0.5%)作比较建立金黄地鼠颊囊癌模型,采用组织学形态分析两种方法在癌变过程中不同病理阶段的进行大体观察、监测体重并计算肿瘤生长率异同,筛选合适的注射浓度建立口腔鳞癌动物模型。
2.应用碳离子束辐照荷颊囊癌金黄地鼠,观测0、4、6、8、12Gy剂量的重离子束对荷颊囊癌金黄地鼠血中微量元素含量的影响,应用原子吸收光谱仪火焰法测定血清中铁(Fe),铜(Cu),锌(Zn),镁(Mg),钙(Ca)5种微量元素的含量。
3.采用免疫组化SP法检测40例荷金黄地鼠颊囊癌组织中Survivin和Caspase-3的蛋白表达,探讨重离子束辐照对肿瘤组织的影响。
结果:1.注射浓度组0.4%、0.3%、0.2%、0.1%分别在13W,9W,12W,16W时出现约为0.8×0.6cm×0.5cm大小的肿瘤;12W时成癌率分别为25%、87%、42%、58%;而0.5% DMBA丙酮涂抹组,12W时成癌率为48%,死亡率为37%,至18W时颊粘膜才出现高分化鳞癌。
2.金黄地鼠成瘤后,血清中微量Fe、Zn、Cu、Mg元素含量均明显低于正常组(P<0.05);再经过不同剂量的重离子束辐照后1月,血清中五种微量元素在低剂量时均呈现下降趋势,随之出现升高,到8Gy再呈降低的趋势,存在一定的剂量-反应关系。
3. Survivin在金黄地鼠口腔颊鳞癌组织(OSCC)中阳性表达28例(28/40),在正常口腔粘膜(NOM)中为零表达(0/8);Caspase-3在金黄地鼠口腔颊鳞癌组织(OSCC)中阳性表达30例(30/40),在正常口腔粘膜(NOM)中为100%(8/8);经统计学分析,Survivin和Caspase-3在OSCC中的表达有统计学意义(P<0.05),(12)~C~(6+)离子辐照对OSCC中Survivin的表达有统计学意义(P<0.05),Caspase-3的表达无统计学意义(P>0.05),辐照与肿瘤临床分期间无统计学意义(P<0.05)。
结论:1.经筛选采用0.3%DMBA丙酮液注射法可诱发形成口腔鳞癌动物模型,缩短实验周期,降低建模成本。
2.经不同剂量的重离子束局部辐照荷瘤金黄地鼠,会影响血清中微量元素的含量,具有一定临床意义。
3.(12)~C~(6+)离子照射对口腔鳞癌组织中的Survivin和Caspase-3蛋白表达产生影响,下调Survivin的表达,间接调节Caspase-3的活性,参与凋亡调控过程。
Objective:To establish hamsters buckle pouch carcinogenesis model by DMBA injection method, The tumor process changes were observed dynamically. To study the effects of trace elements and the expression of apoptosis related genes Survivin, Caspase-3 in hamster buckle pouch carcinogenesis model after carbon ion beams irradiation, so as to provide experimental evidence for Heavy Ion Clinical Therapy of Oral Cancer and risk assessment for Space Radiation.
Materials and Methods:1.This study selected 81 healthy hamsters, they were randomly divided into 3 groups, including the control group (6), the painted group (15), the injected group (60) (also points A, B, C, D, E 5 Groups).With the DMBA acetone solution injection group, The concentration were used to set up for 0.5%,0.4%,0.3%,0.2%,and 0.1%.but the concentration in the painted group was fit for 0.5%. to induce buckle pouch carcinogenesis model in Syrian Golden Hamsters. For the general observation of the model, the body weight has been monitored and the tumor growth rate calculated. Using histological analysis of the similarities and differences in two methods with different pathological stages of.the carcinogenesis process The concentration of injection method by the screening to establish the appropriate carcinogenesis model of oral squamous cell carcinoma.
2. Exposed with carbon ion beam irradiation of cheek pouch carcinoma in hamster, Observation 0,4,6,8,12 Gy dosage of heavy ion beams on the bearing hamster cheek pouch carcinoma of trace elements of content in blood. Application of flame atomic absorption spectroscopy determinated to flexion-extension (Fe) radiographs, copper (Cu), zinc (Zn) and magnesium (Mg), calcium (Ca) the content of trace elements in serum.
3. Immunohistochemical SP method was detected the expression of Survivin and Caspase-3 protein of cheek pouch carcinogenesis in 40 cases bearing golden hamster and to explore on the impact of tumor tissue in Carbon ion beams irradiation.
Results:1 The injection concentrations (0.4%,0.3%,0.2%,0.1%) of DMBA acetone solution have induced the tumors. Respectively, at 13W,9W,12W,16W, the average size is about 0.8cm× 0.6cm×0.5cm; the carcinogenesis rates at 12W were respectively 25%,87%,42%,58%; but 0.5% DMBA solution with painted group, the carcinogenesis rate was 48%,the death rate was 37%. Well differentiated cheek squamous cell carcinoma was obtained at 18W.
2 In the hamsters borne tumor, the contents of Fe, Zn, Cu and Mg in experimental groups with cheek pouch carcinoma were significantly lower than normal group(P<0.05). After irradiated by 0, 4,6,8,12Gy heavy ion beams, the 4Gy group showed a tendency downward. When the irradiation dose reached 6Gy, the contents of Fe, Zn increased. When it reached 12Gy, the contents decreased.. While Cu, Ca and Mg contents in the group at 8Gy reached to their highest value, and had become lowering until 12Gy. All of the results showed a dose-reaction relationship (P<0.05).
3 The number of the expression of positive Survivin protein were 28 among 40 cases in oral squamous cell carcinoma, the positive rate was 70%, but there was no positive individual in normal oral mucosa (0/8); The Caspase-3 expression were 30 among 40 cases, the positive rate was 75% in oral squamous cell carcinoma, and was 100% in normal oral mucosa (8/8). By statistical analysis, we have found there are differences between oral squamous cell carcinoma and normal oral mucosa. After 12C6+ ion irradiation with different doses,Survivin expression was statistically significant (P<0.05) compared with the OSCC group, but the differences of Caspase-3 expression did not occurred. The 12C6+ ion irradiation group compared with OSCC group was not statistically significant (P>0.05).
Conclusions:1 The 0.3% concentration of DMBA acetone solution is more suitable for the formation of oral squamous cell carcinoma model in Golden Hamster in the short term. This method is benefit to shorten the feeding cycle, reduce costs.
2 Exposed with different doses of heavy ion beam irradiation, there are some changes of content in trace elements of serum, which are of clinical significance.
3 12C6+ ion irradiation can reduce the expression of Survivin, indirectly raise the activity of Caspase-3. It reveals to us the 12C6+ ion irradiation may play a critical role in the regulation of the process of cell apoptosis.
引文
[1]Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics,2002. CA Cancer J Clin 2005, 55:74-108.
[2]Reid BC, Winn DM, Morse DE, et al. Head and neck in situ carcinoma:incidence, trends, and survival. Oral Oncol,2000,36:414-420.
[3]Warnakulasuriya S. Global epidemiology of oral and oropharyngeal cancer[J]. Oral Oncol 2009,45:309-316.
[4]Funk GF, Karnell LH, Robinson Raze WK, et al. Presentation, treatment, and outcome of oral cavity cancer:a National Cancer Data Base report[J]. Head Neck,2002,24:165-80.
[5]Sankaranarayanan R, Black RJ, Parkin DM. Cancer survival in developing countries.IARC Scientific publication No.145 Lyon. International Agency for Research on cancer,1998.
[6]Scully C, Bagan JV. Recent advances in Oral Oncology. Oral Oncol,2007,43(2):107-15.
[7]Liam CT, Wang HM, Ng SH, et al. Good tumor control and survivals of squalors cell carcinoma of buckle mucosa treated with radical surgery with or without neck dissection in Taiwan[J]. Oral Once,2006,42:800-809.
[8]Lin CS, Jen YM, Cheng MF, et al. Squalors cell carcinoma of the buckle mucosa:an aggressive cancer requiring multimodality treatment[J]. Head Neck,2006,28:150-157.
[9]Ayer SG, Parham SA, Pay PS, et al. Surgical treatment outcomes of localized squalors carcinoma of buckle mucosa[J]. Head Neck,2004,26:897-902.
[10]Lee KH, Veness MJ, Pearl-Larson T, et al. Role of combined modality treatment of buckle mucosa squalors cell carcinoma[J]. Oust Dent,2005,50:108-113.
[11]Diaz EM JrF, Hollinger C, Zuniga ER, et al. Squalors cell carcinoma of the buckle mucosa:one institutions experience with 119 previously untreated patients[J]. Head Neck, 2003,25:267-273.
[12]Pereira MC, Oliveira DT, Landman G. et al. Histologic subtypes of oral squamous cell carcinoma:prognostic relevance[J]. Can Dent Assoc,2007,73(4):339-344.
[13]Jamal A, Siegel R, Ward E, et al. Cancer statistics,2007.CA Cancer J Clint,2007,57:43-66.
[14]Mork J, Lie AK, Glattre E, et al. Human papillomavirus infection as a risk factor for squalors cell carcinoma of the head and neck[J]. N Engl J Med,2001.344:1125-1131.
[15]B.Mognetti. F.Did Carlo.G.N. Berta. Animal models in oral cancer research[J]. Oral Oncology, 2006.42:448-460.
[16]E.Vairaktaris, S.Spyridonidou, V.Papakosta, et al. The hamster model of sequential oral oncogenesis[J]. Oral Oncology,2008,44:315-324.
[17]Scully C, Field JK, Tanzawa H. Genetic aberrations in oral or head and neck squamous cell carcinoma (SCCHN):1.Carcinogen metabolism, DNA repair and cell cycle control[J]. Oral Oncol 2000,36(3):256-263.
[18]Silverman Jr S. Demographics and occurrence of oral and pharyngeal cancers. The outcomes, the trends, the challenge[J]. Am Dent Assoc,2001,132:7S-11S.
[19]Williams HK. Molecular pathogenesisoforal carcinoma[J]. ClinPatrol,2000,53:165-172.
[20]Raju M R. Particle radiotherapy:historical developments and current status Radiat. Res,1996, 145(4):391-407.
[21]Friedberg E C. A brief history of DNA repair field[J]. Cell Res,2008,18(1):3.
[22]丁小凡,陈龙华.重离子治疗肿瘤的进展[J].中华放射医学与防护杂志,2006,26(5):536-539.
[23]WambersieA.The future of high-LET radiation in cancer therapy proceedings of eulima workshop on the potential value of light ion beam therapy. Nice:EUR 12165 EN,1988, 23-84.
[24]肖国青,张红,李强,等.中国科学院近代物理研究所重离子束治癌进展[J].原子核物理评论,2007,24(2):85-88.
[25]Lillis-Hearne PK, Castro JR. Indication for heavy ions-lessons from berkeley.In:Linz U, eds.Ion beams for tumour therapy. New York:Chapman & Hall,1995,133-141.
[26]Kamada T,Tsujii H,Tsuji H,et al. Benefit of carbon ion radiotherapy in the treatment of radioresistant tumors[J]. AIP Conference Proceedings,2003,680:1142-1145.
[27]Tatsuaki K, Naruhiro M, Tadaaki M, et al. Examination of GyE system for HIMAC carbon therapy[J].Int J Radiat Oncol BioPhys,2006,64(2):650-656.
[28]Satoru Y. New carbon therapy project at GHMC [C]. PTCOG2007,46:58.
[29]GuoChuanling, WangJufang, JinXiaodong, et al. Studies on advantages of heavy ions in radiotherapy compared with x rays.Nuelear Instruments and Methods in Physics Research, 2007,259,997-1003.
[30]H.Stelzerand. Heavy Ion Therapy Collaboration.Tumor therapy with heavy ions at GSI. Nuclear Physics B-Prceedings Supplements,1998,61(3):650-657.
[31]Schulz Ertner D, Nikoghosyan A, Thilmann C, et al. Results of carbon ion radiotherapy in 152 patients[J]. Int J Radiat Oncol Biol Phys,2004,58:631-640.
[32]罗光辉,李文建,苏兴桂.重离子与其他放射治疗肿瘤的比较[J].广东医学,2007,28(1):159-161.
[33]李强.人类征服癌魔的利器---重离子束治疗肿瘤综述.科学新闻,2007,11:44-45.
[34]Luwei Zhang, Hong Zhang, Xiaofu Zhang, et al. Assessment of 12C6+ ionirradiation induced biological changes in wheat seedlings[J]. Nuclear Science Techniques,2008,19.
[35]杨琴,刘复兴.论辐照在医疗领域中的应用前景[J].咸宁学院学报,2010,30(10):9-10.
[36]Wilson R R. Radiological use of fast protons [J]. Radiology,1946,47:487.
[37]TbiasI C A, Robert J E, Lawrence J H, et al. Irradiation of hypophysectomy and related studies using 340MeV protons and 190MeV deuterons [J]. Peaceful Uses at Energy,1956,10: 95.
[38]Paretzke H G.. The First International Workshop on Systems Radiation Biology:a New Approach of Solve Old Questions[J]. Radiat Environ Biophys.2008,47(1):3-4.
[39]Kaminski J M; Shinohara E; Summers J B, et al. The Controversial Abscopal Effect[J] Cancer Treat Rev.2005,31(3):159-72.
[40]S.X. Fang, X.L. Guan, J.Y. Tang, et al. APTF a Dedicated Proton Therapy Facility[J]. Chinese Physics C,2010, Vol,34(03):383-388.
[41]闵锐.辐射生物学研究发展的趋势---系统辐射生物学[J].辐射防通讯,2009,29(5):12-17.
[42]周永增.辐射防护的生物学基础---辅射生物效应[J].辐射防护,2003,23(2):90-101.
[43]Th.Stohlker, T.Beicf, H.E Beyer.et al.AtomiePhysicswithhighly-charged heavy ions at the GSI future facility:The seientific Program of the SPARC collaboratinn. Nudear Instruments and Methods in Physics Research Section B:Beam interactions with Materials and Atoms, 2005,235(1-4):494-497.
[44]Guo Chuanling, Wang Jufang, Jin Xiaodong, et al. Studies on advantages of heavy ions in radiotherapy compared with y-rays[J]. Nuclear Instruments and Methods in Physics Research B,2007,259,997-1003.
[45]YFuosawa, K. Fukutsu, H.Itsukaiehi, et al. Biological effectiveness by the different heavy ion species with the same LET proceeding of the second workshop on physical and biological research with heavy ions,1992,31:p11.
[46]Daniel D, StePhen PJ. DNA PK ATM and ATR assensors of DNA damage:variations on a theme? Current Opinion in Cell Biology,2001,13:225-231.
[47]Danielsson A, Karlsson E, Dell EU.The Biological effect of Pentoxifylline on the survival of human head and Neek Caneer Cells treated with continuous low and High Dose rate irradiation[J]. Cancer Res Clin Oncol,2005,131:459-467.
[48]Kerr JFR. Apoptosis:a basic biological phenomenon with wide ranging implication in tissue kinetic[J].BrJCancer,1972,26:239.
[49]C. Lajolo, M. Giuliani, A. Sgambato, E. et al. N-(4-hydroxyphenyl)all-trans-retinamide (4-HPR) high dose effect on DMBA induced hamster oral cancer:a histomorphometric evaluation.[J] Int Oral Maxillofac. Surg.2008,37:1133-1140.
[50]Deepak Kanojia, Milind M. Vaidya.4-Nitroquinoline-l-oxide induced experimental oral carcinogenesis[R]. Oral Oncology,2006,42,655-667.
[51]Seungwon Kim, MD. Animal models of cancer in the head and neck region[R]. Clinical and Experimental Otorhinolaryngology 2009,2(2):55-60.
[52]F. H. White. K. Gohari. Quantitative studies of hemidesmosomes during progressive DMBA carcinogenesis in hamster cheek-pouch mucosa[J]. Br J Cancer,1981,44(3):440-450.
[53]Rikako Suzuki, Hiroyuki Kohno, Masumi Suzui, et al. An animal model for the rapid induction of tongue neoplasms in human c-Ha-ras proto-oncogene transgenic rats by 4-nitroquinoline 1-oxide:its potential use for preclinical chemoprevention studies[J]. Carcinogenesis,2006,27(3):619-630.
[54]SalleyJJ, KreshoverSJ. The effects of topical application of carcinogens on palatal mucosa of the hamster[J]. Oral Surg,1959,12:501.
[55]WHO collaborating centre for oral precancerous lesion Oral Surg,1978,46:158.
[56]Banoczy J, Csiba, A. Occurrence of epithelial dysplasia in oral leukoplakia. Analysis and follow up study of 12 cases. Oral Surg. Oral Med. Oral Pathol,1976,42,466-774.
[57]Pindborg JJ,Reichart PA,Smith CJ,et al. Histological Typing of Cancer and Precancer of the Oral Mucosa[M].Berlin:WHO, Springer-Verlag,1997,35-38.
[58]周曾同,张水龙,王英,等.金地鼠烦囊上皮异常增生12项病理特征的光镜分析[J]上海口腔医学,1997,6(1):32-35
[59]Dipple A, Pigott MA, Bigger AH, et al.7,12-Dimethylbenz(a)anthracene-DNA binding in mouse skinresponse of different mouse strainsand effect sofvarious modifiers of carcinogenesis[J]. Carcinogenesis,1984,5(8):1087-1090.
[60]Dipple A, Piggot M. Moschel RC,et al. Evidence that binding of 7,12-dimethylbenz-(a)anthracene to DNA in mouse embryo cell cultures results in extensive substitution of both adenine and guanine residues[J]. Can Res,1983,43:4132-4135.
[61]E.Vairaktaris, S.Spyridonidou, V.Papakosta, et al. The hamster model of sequential oral oncogenesis[J]. Oral Oncology,2008,44:315-324.
[62]SUN Rui, ZHANG Jean-gang, GUO Chuan-bin. Establishment of cervical lymph node metastasis model of squalors cell carcinoma in the oral cavity in mice[J]. Chinese Medical Journal,2008,121 (19):1891-1895.
[63]Cao X, Liu X, Hao Z, et al. Establishment of rat submandibular gland squamous cell carcinoma induced by DMBA[J]. Hua Xi Kou Qiang Yi Xue Za Zhi,2000,18(3):156-158.
[64]C. Lajolo, M. Giuliani, A. Sgambato,et al. N-(4-hydroxyphenyl)all-trans-retinamide (4-HPR) high dose effect on DMBA induced hamster oral cancer:a histomorphometric evaluation[J]. Int.Oral Maxillofac. Surg,2008,37:1133-1140.
[65]Morris AL. Factors influencing experimental carcinogenesis in the hamster cheek pouch[J]. Dent Res,1961,40:3-15.
[66]宋宇峰,温玉明.二甲基苯比蒽诱导金黄地鼠颊囊癌的模型建立及组织学研究[J].华西口腔医学,1994,2(2):111-113.
[67]李永生,杨燕DMBA诱发金黄地鼠颊囊癌的实验研究[J].中国癌症杂志,2003,13(2):150-152.
[68]Avila G, Aguilar L, Benitez S, et al. A.Inflammatory responses in the intestinal mucosa of gerbils and hamsters experimentally infected with the adult stage of Taeniasolium[J]. International Journal for Parasitology,2002,32(10):1301-1308.
[71]钟南保.重离子治疗肿瘤进展[R].福州总医院学报,2006,13(2):116-118.
[72]黄秋婵,韦友欢,石景芳.微量元素锌对人体健康的生理效应及其防治途径[J]. Studies of Trace Elements and Health,2009,26(1):.
[73]陆艳琦.微量元素与人体健康[J].河南教育学院学报,2002,11(4):36.
[74]伍明江,张俊巍,蒋朝晖.辐射损伤与微量元素[J]. Studies of Trace Elements and Health. 2006,23(2):51.
[75]单振芬.微量元素检测的方法学讨论[J]. Studies of Trace Elements and Health, 2009,26(6):63-64.
[76]Kaminski P, Kurhalyuk N, Kasprzak M, et al. The impact of element-element interactions on antioxidant enzymatic activity in the blood of white stork (Ciconia ciconia) chicksfJ]. Arch Environ Contam Toxicol,2009,56(2):325-337.
[77]高永.电离辐射对小鼠血清微量元素的影响[J]. Chinese Journal of Public Health,1996. 15(2):94.
[78]罗文海.γ射线对小鼠血清微量元素含量的影响[J].Chinese Journal of Public Health,2003, 19(10):1212.
[79]邓春富,杨彦吕,孙长伏.60~Co-γ射线照射犬下颌骨对血清中微量元素的影响[J].现代口腔医学杂志,1999,13(2):128.
[80]Maksimova, I A Klimenko, L L Protasova,0 V. Macro and micro element balance in tissue of different organs of experimental animals during their physiological and accelerat radiationaging [J]. AdvGerontol,2008,21(2):218-225.
[81]方允中.γ射线离体照射对铜锌超氧化物歧化酶的理化性质的影响[J].科学通报,1984.15,943-946.
[82]Shkala LV. Correction of trace element metabolism in patients with breast cancer during radiation therapy[J]. Vopr Onkol,1997,43(3):273.
[83]侯玮玮,刘斌,张格祥等.重离子辐照对小鼠血清微量元素含量的影响[J].现代预防医学,2008,35(16):3060.
[84]刘金环.微量元素与辐射损伤[J].微量元素与健康研究,2003,20(3):45-46.
[85]Garibov RE, Kniazeva GV, Lukianova SN, et al. Level of microelements-metals in animal blood as an informative indicator of response to low-intensity short duration electromagnetic irradiation Radiats Biol Radioecol.2005,45(5):587.
[86]Zagorskia NG, Kudiasheva AG, Shevchenko OG, et al. Influence of the chronic gamma-irradiation at the low doses during the near ontogenesis periods on the oxidative processes in rodent organs [J]. Radiats Biol Radioecol,2007,47(4):493.
[87]郭传玲,干菊芳,魏巍等.高LET重离子照射人肿瘤细胞的DNA双链断裂及修复研究[J].核技术,2007,30(9):752-753.
[88]王菊芳,李文建,周光明等.三种瘤细胞对重离子和γ射线的辐射敏感性[J].肿瘤2001,21(4):249-250.
[89]Poter AG, Janicke RU. Emerging roles of caspase-3 in apoptosis[J]. Cell Death Differ,1999, 6(2):99-104.
[90]Michael J. Duffy, Norma O'Donovan, Donal J. Brennan, et al. Survivin:A promising tumor biomarker[J]. Cancer Letters,2007,249:49-60.
[91]Tam m I, Wang Y, Sausville E, et al. IAP family protein survivin inhibits caspases and anticancer drugs[J]. Cancer Res,1998,58:5315-5320.
[92]Kawasaki H, Altieri DC, Lu CD, et al. Inhibition of apoptosis by survivin predicts shorter survival rates in colorectal cancer[J].CaneerRes,1998,58(22):5071-5074.
[93]Adida C, Berrebi D, Peuehmaur M, et al. Anti-apoptosis gene surviving and prognosis of neuroblastoma [J]. Laneet,1998:882-883.
[94]Ambrosini G, Adida C, Sirugo G, et al. Induction of apoptosis and inhibition of cell proliferation by Survivin gene targeting[J]. Biol Chem,1998,273:11177-11182
[95]田晓艳,其木格,苏日娜Survivin与Caspase-3在妊娠期高血压疾病患者胎盘组织中的表达及意义[J].内蒙古医学杂志,2010,42(11):1288-1291.
[96]Suzuki A, Ito T, Kawano H, et al. Survivin Initiates procapase3/p21 complex formation as a result of interaction with CDK4 to resist Fas mediated cell death[J]. Oncogene,2000, 19(10):1346-1353.
[97]O Connor DS, Grossman D, Pleseia J, et al. Regulation of apoptosis at cell division by p34 cdc2 phosphorylation of survivin[J]. Proc Nat lAcad SciUSA,2000,13103-13107
[98]Dario C. Altieri. The molecular basis and potential role of survivin in cancer diagnosis and therapy [J]. Trends in Molecular Medicine,2001,7(12):542-547.
[99]Miller L. Anexegesis of APs:salvation and surprises from BIR motifs[J]. Trends Cell Biol, 1999,9(8):323-328
[100]Grossman D, M cNiff M, Li F, et al. Expression of the apoptosis inhibitors surviving in normelanomaskin cancer and gene targeting in a keratinocyte cell line[J]. Lab Invest,1999, 79(9):1121-1126.
[101]潘涛Survivin和Caspase-3在口腔鳞状细胞癌中的表达及其意义.硕十论文.2005,p19.
[102]LoMuzio L, Pannone G, Staibano S, et al. Survivin expression in oral squalors cell careinoma[J]. Br J Caneer.2003,89(12):2244-2248.
[103]高文信,刘洪静,王颖等.Caspase-3在口腔癌前病变及鳞状细胞癌中的表达研究[J].现代口腔医学杂志,2003.17(2):109-111.
[104]Maryla K, Hongwang, Stanislwa K, et al. Inununohisto chemical analysis of in vivo patterns of expression of CPP32(Caspase-3), a cell death Protease[J]. Cancer Res,1997, 57:1605-1613.
[105]蒋月,葛红,叶柯等Survivin和Caspase-3在食管鳞癌术前放疗前后的表达与预后关系分析[J].中国肿瘤临床与康复,2010,17(6):481-483.
[106]汪洋,施学辉.肿瘤潜在倍增时间的基础理论和临床应用研究进展[J].中国癌症杂志,1999,9(2):150-156.
[107]Kumkum Jha, Mridula Shukla, Manoj Pandey. Survivin expression and targeting in breast cancer[J]. Surgical Oncology,2011,1-7.
[108]Xiaoyuan C, Longbang C, Jinghua W, et al. Survivin:a potential prognostic marker and chemoradiotherapeutic target for colorectal cancer[J]. Ir J Med Sci,2010,179(3):327-335.
[109]Farnebo L, Jerhammar F, Ceder R, et al. Combining factors on protein and gene level to predict radioresponse in head and neck cancer cell lines[J]. Oral Pathol Med.2011.
[110]Gino Marioni, Emiliano D'Alessandro, et al. Survivin multifaceted activity in head and neck carcinoma:Current evidence and future therapeutic challenges[J]. Acta Otolaryngol.2010, 130(1):4-9.
[111]Zakir Khan, Noor Khan, Ram P, et al. Down-regulation of survivin by oxaliplatin diminishes radioresistance of head and neck squamous carcinoma cells[J]. Radiotherapy and Oncology, 2010,96,267-273.
[112]Su Liping, Wang Ying, Xiao Mingzhen, et al. Up-regulation of survivin in oral squamous cell carcinoma correlates with poor prognosis and chemoresistance[J]. Oral and maxillofacial pathology,2010,110(4):484-491.
[1]Scully C, Field JK, Tanzawa H. Genetic aberrations in oral or head and neck squamous cell carcinoma (SCCHN):.Carcinogen metabolism, DNA repair and cell cycle control[J]. Oral Oncol 2000,36(3):256-63.
[2]Sanjoy KD. Fate of DMBA in the mouse mammary gland during intiation and promotion stages of carcinogenesis in vitro[J].Cancer Res,1989;49:920.
[3]B.Mognetti, F.Did Carlo, G.N. Berta. Animal models in oral cancer research[J].Oral Oncology, 2006,42:448-460.
[4]Verde M, Carom N, Dean D.4NQO oral carcinogenesis:animal models, molecular markers and future expectations[J]. Oral Once,2005,41:337-9.
[5]AndrogenicS,SavoryJ F,FrontletC,Moniker,et al.7,12-Dimethyl(a)benzanthracene induced early squamous cecarcinoma in Golden Syrian hamsters:evaluation of an animal model and comparison with early forms of human squalors cell carcinoma in the upper aero digestive tract[J].Into J Patrol,1996,77:7-14.
[6]Steindler NE, Reader PC. Experimental induction of oral squalors cell carcinomas in mice with 4-nitroquinolone-1-oxide[J]. Oral Surge Oral Med Oral Patrol,1984,57:524-31.
[7]Deepak Kanojia, Melinda M.Vida.4-Nitroquinoline-l-oxide induced experimental oral carcinogenesis[J].Oral Oncology,2006,42:655-667.
[8]Brachium BJ, Leemans CR. Brinkerhoff RH. A genetic progression model of oral cancer: current evidence and clinical implications[J].OralPathol Med,2004,33:p22.
[9]Williams HK. Molecular pathogenesis of oral carcinoma[J].Clin Patrol,2000,53:165-72.
[10]Dipple A, Pigott MA, Bigger AH, et al.7,12-Dimethylbenz(a)anthracene-DNA binding in mouse skinresponse of different mouse strainsand effect sofvarious modifiers of carcinogenesis[J]. Carcinogenesis 1984,5(8):1087-90.
[11]Dipple A, Piggot M, Moschel RC,et. al. Evidence that binding of 7,12-dimethylbenz(a)
anthracene to DNA in mouse embryo cell cultures results in extensive substitution of both adenine and guanine residues[J]. Can Res,1983,43:4132-4135.
[12]Shklar G, Eisenberg E, Flynn E. Immunoenhancing agents and experimental leukoplakia and carcinoma of the hamster buccal pouch[J]. Prog Exp Tumour Res,1979,24:1269-82.
[13]Avila G, Aguilar L, Benitez S, et.al. Inflammatory responses in the intestinal mucosa of gerbils and hamsters experimentally infected with the adult stage of Taeniasolium[J]. International Journal for Parasitology,2002,32(10):1301-1308.
[14]Lin CS, Jen YM, Cheng MF, et al. Squalors cell carcinoma of the buckle mucosa:an aggressive cancer requiring multimodality treatment[J]. Head Neck,2006,28:150-7.
[15]Hawkins BL, Heniford BW. Ackermann M. et al.4NQO carcinogenesis:a mouse model of oral cavity squalors cell carcinoma[J].Head Neck,1994.16(5):424-432.
[16]Tania J. Shish H.Hiai H, et al. Quantitative trait locate affecting 4-nitroquinoline 1-oxide-induced tongue carcinogenesis in the rat[J]. Cancer Res,1998,58:1660-1664.
[17]Yabushita H. Effects of electro chemotherapy on Cask cells derived from a cervical squalors cell carcinoma[J].Gynecol Oncol.1997,65:297-303.
[18]S.Moure, Y. S. Oath, X. LL H et al. Rapid tumor necrosis induced by electro chemotherapy with intratumoral injection of bloodying in a hamster tongue cancer model.lent[J].Oral Maxillofacial.Surge.2000,29:119-125.
[19]Fujita K, Kaku T, Sasaki M, et al. Experimental production of lingual carcinoma in hamsters by local application of 9,10-dimethyl-1,2-benzanthracene[J].Dent lies,1973,52:327-32.
[20]Mori M, Miyaji T, Murata I, et al. Histochemical observations on enzymatic processes of experimentalcarcinogenesis in hamster cheek pouch[J]. Cancer Res 1962,22:1323-1326.
[21]Solt DB. Localization of gamma-glutamyl transpeptidase in hamster buccal pouch epithelium treated with 7,12-dimethylbenzaanthracene[J]. Natl Cancer Inst 1981,67:193-200.
[22]Tang XH, Knudsen B, Bemis D, et al. Oral cavity and esophageal carcinogenesis modelled in carcinogen-treated mice[J]. Clin Cancer Res,2004,10:301-13.
[23]Wong PN, Wilson DF.4-Nitroquinoline-l-oxide-induced carcinogenesis in the rat palate[J].Oral Pathol 1983,12:375-84.
[24]Sood S,Schiff SJ,Yang CS,et al. Selection of topically applied non-steroidal anti-inflammatory drugs for oral cancer chemoprevention[J].Oral Oncol,2005,41:562-567.
[25]Amok Dub, Subhead Sharma, Praveen Kumar Gupta. Evaluation of chlorine p6 for photodynamic treatment of squalors cell carcinoma in the hamster cheek pouch model[J].Oral Oncology,2006,42:77-78.
[26]Zhou ZT, Yang Y, Ge JP. The preventive effect of salvianolic acid B on malignant transformation of DMBA-induced oral premalignant lesion in hamsters[J]. Carcinogenesis,2006,27:826-832.
[27]Yang Y, Zhou ZT, Ge JP. Effect of Einstein on DMBA-induced oral carcinogenesis in hamster[J]. Carcinogenesis,2006,27:578-583.
[28]Salley JJ. Experimental carcinogenesis in the cheek pouch of the Syrian hamster[J].Dent Res 1954,33:253-62.
[29]SalleyJJ, KreshoverSJ. The effects of topical application of carcinogens on palatal mucosa of the hamster[J].Oral Surg,1959,12:501.
[30]Seungwon Kim MD. Animal models of cancer in the head and neck region[J]. Clinical and Experimental Otorhinolaryngology,2009,2(2):55-60.
[31]郑金华.二甲基苯并蒽涂抹联合创伤诱发舌癌模型的比较研究[R]. Anat Research,2007, 29(6):405-409.
[32]Dipple A, Piggot M, Moschel RC.et al. Evidence that binding of 7,12-dimethylbenz-(a)anthracene to DNA in mouse embryo cell cultures results in extensive substitution of both adenine and guanine residues[J]. Can Res,1983,43:4132-4135.
[33]E.Vairaktaris. S.Spyridonidou, V.Papakosta, et al. The hamster model of sequential oral oncogenesis[J]. Oral Oncology,2008,44:315-324.
[34]Bindles EM, Vanden Brekel MW. Development of a conditional mouse model for head and neck squamous cell carcinoma[J].Adv Otorhinolaryngol.2005,62:1-11.
[35]TakeshNomura,Takahikhibahara,AkiraKatakura, et.al. Establishment of a marine model of bone invasion by oral squamous cell carcinoma[J]. Oral Oncology,2007,43:257-262.
[36]Yang M. Jiang P, Sun FX, et al. A fluorescent orthotopic bone metastasis model of human prostate cancer[J]. Cancer Res.1999,59:781-6.
[37]Rosol TJ, Tannehill-Gregg SH, LeRoy BE, et al. Animal models of bone metastasis[J]. Cancer, 2003.97:748-57.
[38]Odukoya O, Schwartz J, Weichselbaum R, et al. An epidermoi carcinoma cell in ederived from hamster7,12-dimethyl-benzaanthracene-induced buccal pouch tumors[J]. Natl Cancer Inst 1983,71:1253-64.
[39]Meng CL, Shklar G, Albright J. A transplantable anaplastic oral cancer model[J]. Oral Surg Oral Med Oral Pathol,1982,53:179-89.
[40]Morifuji M, Taniguchi S, Sakai H, et al.Differentialexpression of cytokeratin after orthotopic implantation of newly established human tongue cancer cell lines of defined metastaticability[J].
AmJ Pathol.2000,156(4):1317-1326.
[41]Xia-cheng Liu, Li-qun Xu, Li-xin Sui, et al. Site-specific lymphatic mapping relative to lingual septum in localizing the regional lymph nodes of tongue-an animal study[J]. Surgical Oncology,2009, xx:1-9.
[42]Goldstein BD. Cancer and Environment[J]. Science,1992,255,5074:904.
[43]Wall RJ, Seidel GJr. Transgenicfarmanimal[R]. Theriogenology,1992,38:337.
[44]ChalbergW, VankovA, Molnar FE, et al. Gene transfer to rabbit retina with electron avalanche transfection[J]. Invest Ophthalmol Vis Sci,2006,47:408.
[45]Hammet RE, Puresl VG, Rexroad CE, et al. Production of transgenic rabbits, sheep and pigs by microinjection[J]. Nature,1985,315:680.
[46]Raimondi AR, Molinolo A, Gutkind JS. Rapamycin prevents early onset of tumorigenesis in an oral-specific K-ras and p53 two hit carcinogenesis model[J]. Cancer Res,2009,69(10): 4159-4166.
[47]Moral M, Segrelles C, Lara MF, et al. Akt activation synergizes with Trp53 loss in oral epithelium to produce a novel mouse model for head and neck squalors cell carcinoma[J]. CancerRes,2009,69(3):1099-1108.