MDM2蛋白和Rb蛋白在喉鳞状细胞癌中的表达及其相关性的研究
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摘要
目的:喉癌(carcinoma of the larynx)是头颈部常见的恶性肿瘤,约占头颈肿瘤的13.9%,占全身恶性肿瘤的2.1%。喉部恶性肿瘤中96%～98%为鳞状细胞癌。近年来喉癌的发病率有明显增加的趋势。喉鳞状细胞癌的病因迄今尚未完全明了,目前认为与性别,吸烟,空气污染,喉白斑病等有关,但现在越来越多的认为与基因表达改变有关。
癌基因MDM2是一种进化保守基因,MDM2首先在自发转化的BALB/C细胞中发生基因扩增被发现。Rb蛋白是从视网膜母细胞瘤中发现并克隆出的第一个人类肿瘤抑制基因,在对细胞生长的调控中,Rb蛋白的去磷酸化状态是维持细胞在G0或G1期的活化形式,抑制和控制细胞生长和增殖,而磷酸化的Rb蛋白将细胞由这种控制中释放出来,使细胞进入DNA合成及有丝分裂期。SV40LT Ag与其它癌基因在DNA病毒的复制与转化中起重要作用,他们与Rb蛋白的结合,使人们对这些能与肿瘤抑制蛋白相结合的癌蛋白的真核细胞同源性发生了兴趣, MDM2蛋白同SV40LT Ag相似,可以抑制Rb的转录调节功能。但是目前对于MDM2与Rb之间的关系还不清楚。
本实验主要研究MDM2蛋白和Rb蛋白在喉鳞状细胞癌组织中的表达情况,通过观察MDM2蛋白和Rb蛋白在喉鳞癌,癌前病变,声带息肉,癌旁正常喉粘膜组织中的表达,借以初步探讨其肿瘤特异性及其与喉鳞癌发生、发展的相关性,并探寻喉癌早期诊断、肿瘤病理分级及其临床分期评估的参考指标,为临床制定治疗方案提供分子生物学参考依据。
方法:选择河北医科大学第二医院耳鼻喉科2008—2009年有完整临床病例资料,均经病理检查证实的手术切除标本共70例,喉鳞状细胞癌组40例,癌周正常组织组取10例(距肿瘤>0.5cm),声带息肉组10例,癌前病变组10例(成人型喉乳头状瘤10例)。其中喉鳞状细胞癌组40例患者均为男性,年龄54—71岁,平均62.57岁,术前均未接受放疗和化疗,经病理证实均为喉鳞状细胞癌组织,按2002年国际抗癌联盟(UICC)标准进行临床特征分期,其中Ⅰ期5例,Ⅱ期7例,Ⅲ期16例,Ⅳ12例;按肿瘤分化程度将其分级,其中高分化18例,中分化12例,低分化10例。每例手术标本取材后即放入10%的福尔马林固定液,常规石蜡包埋,以5μm厚度连续切片,石蜡切片经常规二甲苯脱蜡,梯度酒精至水,去除内源性过氧化物酶,微波抗原修复,SP法分别行MDM2和RB免疫组织化学染色检测。DAB显色,苏木精复染,切片脱水、透明、树胶封片。
统计学处理:所有实验数据均用SPSS17.0统计软件进行分析,数据比较采用X2检验, MDM2蛋白和Rb蛋白的相关性采用Spearman等级相关分析,以P<0.05为差异有统计学意义。
结果:MDM2蛋白的阳性表达在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌组之间均有显著性差异,呈升高趋势。
Rb蛋白的阳性表达在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌组之间均有显著性差异,呈降低趋势。
MDM2蛋白的阳性表达在高分化、中分化、低分化喉鳞癌之间有显著性差异,呈升高趋势。
RB蛋白的阳性表达在高分化、中分化、低分化喉鳞癌之间有显著性差异,呈降低趋势。
MDM2蛋白的阳性表达在Ⅰ、Ⅱ、Ⅲ、Ⅳ期喉鳞癌间有显著性差异,呈升高趋势。
Rb蛋白的阳性表达在Ⅰ、Ⅱ、Ⅲ、Ⅳ期喉鳞癌间有显著性差异,呈降低趋势。
MDM2蛋白的过度表达和Rb蛋白的失活和喉癌的发展密切相关,呈负相关性。
结论:MDM2蛋白在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌中的阳性表达升高,Rb蛋白在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌中的阳性表达降低,二者在喉鳞癌的表达具有负相关性,因此二者相结合可作为喉癌早期诊断的指标。
MDM2蛋白在喉鳞癌病理分级高分化、中分化、低分化的阳性表达和在临床特征分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期的阳性表达依次升高,Rb蛋白在喉鳞癌病理分级高分化、中分化、低分化的阳性表达和在临床特征分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期的阳性表达依次降低,通过检测喉鳞癌中MDM2蛋白和Rb蛋白的阳性表达可以作为评估肿瘤病理分级及其临床分期评估的参考指标,为临床制定治疗方案提供分子生物学参考依据。
Objective : Laryngocarcinoma is in high morbidity of head and neck malignancies. It accounts for 13.9% of head and neck tumours and represents 2.1% of all malignancies. 96%-98% of the laryngeal malignant tumor is squamous cell carcinoma. The morbidity of the laryngocarcinoma progressed obviously in recent years. The causes of laryngeal squamous cell carcinoma have not yet fully understood. People think it has a relationship with gender, smoking, air pollution, leukoplakia of larynx and other factors. But more and more people related it to the changes of the gene expression nowadays.
MDM2 is an evolutionarily conserved gene. It first be found in the BALB/C cells which are in spontaneous transformation when gene amplification was performed. Rb is the first tumor suppressor gene of the human being which was found in retinoblastoma cell and be cloned. In the cell growth regulation, the dephosphorylation form of the Rb protein can maintain the activation of the cells in G0 or G1 phase, suppress and control cell growth and proliferation, while the phosphorylation form released from this control and make the cells to enter to the DNA synthesis and mitotic phase. SV40LT Ag and other cancer genes play an important role in DNA replication and transformation of the virus. Because their combination with the Rb protein, people interested in the homology of the eukaryotic cells in which tumor suppressor protein can make a combination with the oncoprotein. MDM2 protein, similar to SV40LT Ag, can inhibit the transcription and regulatory function of Rb. But the relationship between MDM2 and Rb is unclear at present.
Major study in this experiment is the expression of MDM2 protein and Rb protein in laryngeal squamous cell carcinoma. By observing the expression of MDM2 protein and Rb protein in laryngeal squamous cell carcinoma, precancerous lesions, vocal cord polyps, peritumoral normal laryngeal mucosa tissues, we investigate its specificity and relationship with the occurrence and development of the laryngeal squamous cell carcinoma as well as to find a reference index for the early diagnosis of laryngeal cancer ,pathological and clinical staging evaluation .This will provide molecular biology reference for the clinical treatment planning.
Methods: 70 cases were choosed from 2008-2009 ENT of The Second Affiliated Hospital of Hebei Medical University with complete clinical data and be confirmed by pathological examination including 40 cases of laryngeal squamous cell carcinoma ,10 cases of peritumoral normal tissue group(from the edge of the tumor > 1.5cm),10 cases of vocal cord polyp group and 10 cases of precancerous lesion group(10 cases of adult laryngeal papilloma ). All the 40 laryngeal squamous cell carcinoma patients were male, aged from 54-71, have a mean age of 62.57. They all had no preoperative radiotherapy and chemotherapy and were pathologically confirmed as laryngeal squamous cell carcinoma. According to the International Anti - Cancer Coalition (UICC) 2002 standard which was separated by its clinical features,5 cases of stageⅠ,7 cases of stageⅡ, 16 cases of stageⅢ, 12 cases of stageⅣwere included. According to the degree graded by tumor differentiation,18 cases of well differentiated laryngeal squamous cell carcinoma, 12 cases of moderately differentiated laryngeal squamous cell carcinoma, 10 cases of poorly differentiated laryngeal squamous cell carcinoma were included. Each specimen was put into the 10% formalin fixative immediately after its surgical remove, conventional paraffin-embedding, serial sections by 5μm thick, deparaffinaged by xylene in convention, dehydrated by alcohol in gradient, endogenous peroxidase removed ,antigen repaired by microwave, MDM2 and RB immunohistochemical staining detected by SP method, DAB stain, hematoxylin stain, dehydrated, transparented ,mounted by gum.
Statistical analysis: All experimental datas were analyzed using statistical software SPSS17.0, compared by X 2 test. Spearman rank correlation analysis was used to find the correlation between MDM2 protein and Rb protein, P <0.05 is statistically significant.
Results: The positive expression of MDM2 protein in the peritumoral normal tissue, vocal cord polyps, laryngeal precancerous lesions and laryngeal squamous cell carcinoma have significant difference, an upgrade trend was showed.
The positive expression of Rb protein in peritumoral normal tissue, vocal cord polyps, laryngeal precancerous lesions and laryngeal squamous cell carcinoma have significant difference, a degrade trend was showed.
The positive expression of MDM2 protein in well differentiated、moderately differentiated laryngeal cancer、poorly differentiated laryngeal squamous cell carcinoma has significant difference,showed an upgrade trend.
The positive expression of Rb protein in well differentiated laryngeal squamous cell carcinoma、moderately differentiated laryngeal squamous cell carcinoma、poorly differentiated laryngeal squamous cell carcinoma has significant difference,showed a degrade trend.
The positive expression of MDM2 protein in theⅠ,Ⅱ,Ⅲ,Ⅳstage of laryngeal squamous cell carcinoma has significant difference, showed an upgrade trend.
The positive expression of Rb protein in theⅠ,Ⅱ,Ⅲ,Ⅳstage of laryngeal squamous cell carcinoma has significant difference, showed a degrade trend.
The over expression of MDM2 protein and the inactivation of Rb protein is closely related to the development of laryngeal cancer, showing negative correlation.
Conclusion: The positive expression of MDM2 protein in peritumoral normal tissue、vocal cord polyps、laryngeal precancerous lesions and laryngeal squamous cell carcinoma was increased while the positive expression of Rb protein in peritumoral normal tissue、vocal cord polyps、laryngeal precancerous lesions and laryngeal squamous cell carcinoma was decreased. This two have a negative correlation in the expression of the laryngeal squamous cell carcinoma .So a combination of both can be used as indicators for early diagnosis of laryngeal carcinoma.
The positive expression of MDM2 protein in laryngeal squamous cell carcinoma in well differentiated laryngeal squamous cell carcinoma, moderately differentiated laryngeal squamous cell carcinoma, poorly differentiated laryngeal squamous cell carcinoma which are graded by pathology and in all theⅠ,Ⅱ,Ⅲ,Ⅳstages which are graded by clinical features are increased by order while the positive expression of Rb protein in laryngeal squamous cell carcinoma in well differentiated laryngeal squamous cell carcinoma, moderately differentiated laryngeal squamous cell carcinoma, poorly differentiated laryngeal squamous cell carcinoma and in all theⅠ,Ⅱ,Ⅲ,Ⅳstages are decreased by order. The detection of the MDM2 and Rb protein expression in the laryngeal squamous cell carcinoma can be used as a reference index for the assessment of the pathological grade and the clinical stage as well as to provide molecular biology reference for the clinical treatment planning.
癌基因MDM2是一种进化保守基因,MDM2首先在自发转化的BALB/C细胞中发生基因扩增被发现。Rb蛋白是从视网膜母细胞瘤中发现并克隆出的第一个人类肿瘤抑制基因,在对细胞生长的调控中,Rb蛋白的去磷酸化状态是维持细胞在G0或G1期的活化形式,抑制和控制细胞生长和增殖,而磷酸化的Rb蛋白将细胞由这种控制中释放出来,使细胞进入DNA合成及有丝分裂期。SV40LT Ag与其它癌基因在DNA病毒的复制与转化中起重要作用,他们与Rb蛋白的结合,使人们对这些能与肿瘤抑制蛋白相结合的癌蛋白的真核细胞同源性发生了兴趣, MDM2蛋白同SV40LT Ag相似,可以抑制Rb的转录调节功能。但是目前对于MDM2与Rb之间的关系还不清楚。
本实验主要研究MDM2蛋白和Rb蛋白在喉鳞状细胞癌组织中的表达情况,通过观察MDM2蛋白和Rb蛋白在喉鳞癌,癌前病变,声带息肉,癌旁正常喉粘膜组织中的表达,借以初步探讨其肿瘤特异性及其与喉鳞癌发生、发展的相关性,并探寻喉癌早期诊断、肿瘤病理分级及其临床分期评估的参考指标,为临床制定治疗方案提供分子生物学参考依据。
方法:选择河北医科大学第二医院耳鼻喉科2008—2009年有完整临床病例资料,均经病理检查证实的手术切除标本共70例,喉鳞状细胞癌组40例,癌周正常组织组取10例(距肿瘤>0.5cm),声带息肉组10例,癌前病变组10例(成人型喉乳头状瘤10例)。其中喉鳞状细胞癌组40例患者均为男性,年龄54—71岁,平均62.57岁,术前均未接受放疗和化疗,经病理证实均为喉鳞状细胞癌组织,按2002年国际抗癌联盟(UICC)标准进行临床特征分期,其中Ⅰ期5例,Ⅱ期7例,Ⅲ期16例,Ⅳ12例;按肿瘤分化程度将其分级,其中高分化18例,中分化12例,低分化10例。每例手术标本取材后即放入10%的福尔马林固定液,常规石蜡包埋,以5μm厚度连续切片,石蜡切片经常规二甲苯脱蜡,梯度酒精至水,去除内源性过氧化物酶,微波抗原修复,SP法分别行MDM2和RB免疫组织化学染色检测。DAB显色,苏木精复染,切片脱水、透明、树胶封片。
统计学处理:所有实验数据均用SPSS17.0统计软件进行分析,数据比较采用X2检验, MDM2蛋白和Rb蛋白的相关性采用Spearman等级相关分析,以P<0.05为差异有统计学意义。
结果:MDM2蛋白的阳性表达在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌组之间均有显著性差异,呈升高趋势。
Rb蛋白的阳性表达在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌组之间均有显著性差异,呈降低趋势。
MDM2蛋白的阳性表达在高分化、中分化、低分化喉鳞癌之间有显著性差异,呈升高趋势。
RB蛋白的阳性表达在高分化、中分化、低分化喉鳞癌之间有显著性差异,呈降低趋势。
MDM2蛋白的阳性表达在Ⅰ、Ⅱ、Ⅲ、Ⅳ期喉鳞癌间有显著性差异,呈升高趋势。
Rb蛋白的阳性表达在Ⅰ、Ⅱ、Ⅲ、Ⅳ期喉鳞癌间有显著性差异,呈降低趋势。
MDM2蛋白的过度表达和Rb蛋白的失活和喉癌的发展密切相关,呈负相关性。
结论:MDM2蛋白在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌中的阳性表达升高,Rb蛋白在癌周正常组织、声带息肉、喉癌前病变与喉鳞癌中的阳性表达降低,二者在喉鳞癌的表达具有负相关性,因此二者相结合可作为喉癌早期诊断的指标。
MDM2蛋白在喉鳞癌病理分级高分化、中分化、低分化的阳性表达和在临床特征分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期的阳性表达依次升高,Rb蛋白在喉鳞癌病理分级高分化、中分化、低分化的阳性表达和在临床特征分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期的阳性表达依次降低,通过检测喉鳞癌中MDM2蛋白和Rb蛋白的阳性表达可以作为评估肿瘤病理分级及其临床分期评估的参考指标,为临床制定治疗方案提供分子生物学参考依据。
Objective : Laryngocarcinoma is in high morbidity of head and neck malignancies. It accounts for 13.9% of head and neck tumours and represents 2.1% of all malignancies. 96%-98% of the laryngeal malignant tumor is squamous cell carcinoma. The morbidity of the laryngocarcinoma progressed obviously in recent years. The causes of laryngeal squamous cell carcinoma have not yet fully understood. People think it has a relationship with gender, smoking, air pollution, leukoplakia of larynx and other factors. But more and more people related it to the changes of the gene expression nowadays.
MDM2 is an evolutionarily conserved gene. It first be found in the BALB/C cells which are in spontaneous transformation when gene amplification was performed. Rb is the first tumor suppressor gene of the human being which was found in retinoblastoma cell and be cloned. In the cell growth regulation, the dephosphorylation form of the Rb protein can maintain the activation of the cells in G0 or G1 phase, suppress and control cell growth and proliferation, while the phosphorylation form released from this control and make the cells to enter to the DNA synthesis and mitotic phase. SV40LT Ag and other cancer genes play an important role in DNA replication and transformation of the virus. Because their combination with the Rb protein, people interested in the homology of the eukaryotic cells in which tumor suppressor protein can make a combination with the oncoprotein. MDM2 protein, similar to SV40LT Ag, can inhibit the transcription and regulatory function of Rb. But the relationship between MDM2 and Rb is unclear at present.
Major study in this experiment is the expression of MDM2 protein and Rb protein in laryngeal squamous cell carcinoma. By observing the expression of MDM2 protein and Rb protein in laryngeal squamous cell carcinoma, precancerous lesions, vocal cord polyps, peritumoral normal laryngeal mucosa tissues, we investigate its specificity and relationship with the occurrence and development of the laryngeal squamous cell carcinoma as well as to find a reference index for the early diagnosis of laryngeal cancer ,pathological and clinical staging evaluation .This will provide molecular biology reference for the clinical treatment planning.
Methods: 70 cases were choosed from 2008-2009 ENT of The Second Affiliated Hospital of Hebei Medical University with complete clinical data and be confirmed by pathological examination including 40 cases of laryngeal squamous cell carcinoma ,10 cases of peritumoral normal tissue group(from the edge of the tumor > 1.5cm),10 cases of vocal cord polyp group and 10 cases of precancerous lesion group(10 cases of adult laryngeal papilloma ). All the 40 laryngeal squamous cell carcinoma patients were male, aged from 54-71, have a mean age of 62.57. They all had no preoperative radiotherapy and chemotherapy and were pathologically confirmed as laryngeal squamous cell carcinoma. According to the International Anti - Cancer Coalition (UICC) 2002 standard which was separated by its clinical features,5 cases of stageⅠ,7 cases of stageⅡ, 16 cases of stageⅢ, 12 cases of stageⅣwere included. According to the degree graded by tumor differentiation,18 cases of well differentiated laryngeal squamous cell carcinoma, 12 cases of moderately differentiated laryngeal squamous cell carcinoma, 10 cases of poorly differentiated laryngeal squamous cell carcinoma were included. Each specimen was put into the 10% formalin fixative immediately after its surgical remove, conventional paraffin-embedding, serial sections by 5μm thick, deparaffinaged by xylene in convention, dehydrated by alcohol in gradient, endogenous peroxidase removed ,antigen repaired by microwave, MDM2 and RB immunohistochemical staining detected by SP method, DAB stain, hematoxylin stain, dehydrated, transparented ,mounted by gum.
Statistical analysis: All experimental datas were analyzed using statistical software SPSS17.0, compared by X 2 test. Spearman rank correlation analysis was used to find the correlation between MDM2 protein and Rb protein, P <0.05 is statistically significant.
Results: The positive expression of MDM2 protein in the peritumoral normal tissue, vocal cord polyps, laryngeal precancerous lesions and laryngeal squamous cell carcinoma have significant difference, an upgrade trend was showed.
The positive expression of Rb protein in peritumoral normal tissue, vocal cord polyps, laryngeal precancerous lesions and laryngeal squamous cell carcinoma have significant difference, a degrade trend was showed.
The positive expression of MDM2 protein in well differentiated、moderately differentiated laryngeal cancer、poorly differentiated laryngeal squamous cell carcinoma has significant difference,showed an upgrade trend.
The positive expression of Rb protein in well differentiated laryngeal squamous cell carcinoma、moderately differentiated laryngeal squamous cell carcinoma、poorly differentiated laryngeal squamous cell carcinoma has significant difference,showed a degrade trend.
The positive expression of MDM2 protein in theⅠ,Ⅱ,Ⅲ,Ⅳstage of laryngeal squamous cell carcinoma has significant difference, showed an upgrade trend.
The positive expression of Rb protein in theⅠ,Ⅱ,Ⅲ,Ⅳstage of laryngeal squamous cell carcinoma has significant difference, showed a degrade trend.
The over expression of MDM2 protein and the inactivation of Rb protein is closely related to the development of laryngeal cancer, showing negative correlation.
Conclusion: The positive expression of MDM2 protein in peritumoral normal tissue、vocal cord polyps、laryngeal precancerous lesions and laryngeal squamous cell carcinoma was increased while the positive expression of Rb protein in peritumoral normal tissue、vocal cord polyps、laryngeal precancerous lesions and laryngeal squamous cell carcinoma was decreased. This two have a negative correlation in the expression of the laryngeal squamous cell carcinoma .So a combination of both can be used as indicators for early diagnosis of laryngeal carcinoma.
The positive expression of MDM2 protein in laryngeal squamous cell carcinoma in well differentiated laryngeal squamous cell carcinoma, moderately differentiated laryngeal squamous cell carcinoma, poorly differentiated laryngeal squamous cell carcinoma which are graded by pathology and in all theⅠ,Ⅱ,Ⅲ,Ⅳstages which are graded by clinical features are increased by order while the positive expression of Rb protein in laryngeal squamous cell carcinoma in well differentiated laryngeal squamous cell carcinoma, moderately differentiated laryngeal squamous cell carcinoma, poorly differentiated laryngeal squamous cell carcinoma and in all theⅠ,Ⅱ,Ⅲ,Ⅳstages are decreased by order. The detection of the MDM2 and Rb protein expression in the laryngeal squamous cell carcinoma can be used as a reference index for the assessment of the pathological grade and the clinical stage as well as to provide molecular biology reference for the clinical treatment planning.
引文
1孔维佳,耳鼻咽喉头颈外科学北京;人民卫生出版社,2002.2
2王正敏,陆书昌,现代耳鼻咽喉科学北京;民军医出版社,2001.10
3 Miwa S, Uchida C, Kitagawa K, et al. Mdm22mediated pRB down regulation is involved in carcinogenesis in a p53-independentmanne. Biochem BiophysResCommun, 2006, 340 1 :54 - 61Weiberg R A The retinoblastoma and cell cycle control 1995(2)
4 Yamasaki L. Role of RB tumor suppressor in cancer. Cance TreatRes,2003, 115( 9) :209 - 239宗永生,王连唐,耳鼻咽喉肿瘤病理部分广州;广东科技出版社,2002.5
5黄选兆等,实用耳鼻咽喉头颈外科学北京;人民卫生出版社,2007.12
6 CAHILLY-SNYDER L,YANG-FENG T,FRANCKE U,et al. moccular analysis and chromosomal mapping of amplified genes isolated from a transformed mouse 3T3 cell line. Somet Cell Genet,1987,13(1):235-244
7 OLINER J D,KINZLER K W,MELTZER P S,et al.Amplification of a gene encoding a P53-associated protein in human sarcomas. Nature,1992,358(2): 80-81
8 Weissman AM. Themes and variations on ubiquitylation. Nat Rev Mol Cell Biol,2001,2 3 :169-178
9 Jin Y, Zeng SX, Dai MS, et al. MDM2 inhibits PCAF (p300/CREB-binding protein-associated factor)–mediated P53 acetylation. J Biol Chem, 2002, 277 (34): 30838-30843
10 Yin Y, Luciani MG, Fahraeus. P53 stability and activity is regulated by Mdm2-mediated induction of alternative p53 translation products. Nat Cell Biol,2002,4 6 : 462-467
11李小华,陈德基,江曼,等.实验性肺癌癌变过程中p53,mdm2,K- ras及PCNA蛋白的表达.癌症, 2001; 20(9) : 921– 925
12任义,吴逸明,董海燕,等.肺癌组织中MDM2,P53蛋白的表达.郑州大学学报医学版,2005; 40(1)67– 69
13孙志,细胞周期分子调控与癌变.国外医学遗传学分册,1996;19(1);50
14张晓辉,细胞周期的网络调控系统Cyclins-CDKs-CKIs及其余肿瘤的关系.国外医学肿瘤学分册,1997;25(1):77
15王斌全等,喉癌组织RB基因突变及其与临床分期的关系临床耳鼻咽喉科杂志2003 ,2 17 :89-90
16 Helt AM,Galloway.Mechannisms by which DNA tumor virus oncoproteins target the Rb family of pocket protein.Carcinogenesis, 2003,24(2);159-169
17 Reichelt M, Zang K D, Seifert M, et al. The yeast two-hybrid system reveals no interaction between p73 alpha and SV40 large T-antigen.Arch Virol,1999,144(3):621-626
18 Junen Gershon T, ShifmanO, UngerT, etal. The MDM2 oncoprotein interacts with the cell fate regulator numb.Mol CellBiol, 1998,18:3974-3982
19 Sdek P, Ying H, Chang D L, et al. MDM2 promote sproteasome-dependent ubiquitin-independent degradation of retinoblastoma protein. MolCell, 2005,9(5):699 - 708 .
1王正敏,陆书昌,现代耳鼻咽喉科学北京:人民军医出版社,2001.10
2周梁,董频,临床耳鼻咽喉头颈肿瘤学上海:复旦大学出版社,2008.9
3姜泗长,耳鼻咽喉-头颈外科手术学北京:人民军医出版社,2005.9
4王正敏,临床耳鼻咽喉科学上海:复旦大学出版社, 2007.10
5宗永生,王连唐,耳鼻咽喉肿瘤病理部分广州;广东科技出版社,2002.5
6黄选兆等,实用耳鼻咽喉头颈外科学北京:人民卫生出版社,2007.12
7王家东,耳鼻咽喉头颈肿瘤外科学上海:上海交通大学出版社,2007.9
8王天铎,喉科手术学北京:人民卫生出版社,2007.4
9杨宝琦,耳鼻咽喉科学新进展天津:天津科学技术出版社,2000.3
10罗荣城,肿瘤生物治疗学北京:人民卫生出版社,2006.08
11桑建中等,P53、Rb和P16在喉鳞癌组织中的表达及意义山东医药,2005,45:17-19
12 Reddel RR. The role of the senescence and immortalizationin carcinogenesis. Carcinogenesis, 2000, 21 3 : 477
13 Cook JR,HillDA, Humphrey PA, et al. Squamous cell car cinoma arising in recurrent respiratory papillomatosis with pulmonary involvement: emerging common pattern of clinical features and human papillomavirus serotype association. Mod Pathol,2000,13 8 : 914
14 Rassekh CH, Rady PL, Arany I, et al. Combined Epstein-Barrvirus and human papillomavirus infection in nasopharyngealcarcinoma. Laryngoscope, 1998,108 3 : 362
15 YAMAUCHI T, WATANABE M,KUBOTA T, et al. Cyclooxygenase22 expression as a new marker for patients with colorectal cancer[J ]. Dis Colon Rectum 2002 ,45:98 - 103
16 TSUJII M , KAWANO S, TSUJI S, et al. Cyclooxy2genase regulates angiogenesis induced by colon cancer cells. Cell ,1998 ,93:705 - 716
17 KAKIUCHI Y, TSUJI S, TSUJII M , et al. Cyclooxygenase-2 activity altered the cell-surface carbohydrate antigens on colon cancer cells and enhanced liver me- tastasis. Cancer Res ,2002 ,62:1567 - 1572
18 YUAN A , YU C J , SHUN C T, et al. Total cyclooxygenase-2 mRNA levels correlate with vascular endothelial growth factor mRNA levels , tumor angiogenesis and prognosis in non-small cell lung cancer patients. Int J Cancer ,2005 ,115:545 - 555
19 HARIZI H, JUZAN M , PITARD V , et al. Cyclooxygenase-2-issued prostaglandin enhances the production of endogenous IL-10 , which down-regulates dendritic cell functions. J Immunol ,2002 ,168:2255 - 2263
20 JEONG M A , LEE K W, YOON D Y, et al. Jaceosidin , a pharmacologically active flavone derived from Artemisia argyi , inhibits phorbol2ester2induced upregulation of COX-2 and MMP-9 by blocking phosphorylation of ERK-1 and-2 in cultured human mammary epithelial cells. Ann N Y Acad Sci ,2007 ,1095:458- 466
21陈望燕等,COX-2基因及产物在人类喉癌组织中的表达
22临床耳鼻咽喉头颈外科杂志,2008.6 , 22:532-535
23李晓光,于肖鹏,赵海兰,等. PCNA在口腔鳞癌中的表达及其临床意义.泰山医学院学报,2005,26 1 : 20 - 22
24李正贤等,PCNA、Bcl-2和Bax在喉癌组织中的表达及其意义临床军医杂志,2008.12,36:850-852
25姚军.p27与卵巢癌.华夏医学,2003,16 2 :125-127
26李学忠等,MDM2与p27在下咽癌中的表达及其临床意义临床耳鼻咽喉头颈外科杂志,2004.12 , 18:716-718
27 DONG J T,LAMB PW,RINKER-SCHAEFFER CW,et al.KAI1,a metastasis suppressor gene for prostate cancer on human chromosome 11p11.2 J.Science,1995,268 (5) :8842886
28白伟良,任重,潘子民,等.KAI1基因在喉鳞癌中的表达及其临床意义.中华肿瘤杂志,2005,27( 5):289-291
29 Herrera L, Kakati S, GibasL, et al. Gardner syndrome in a man with an interstitial deletion of 5q. Am J Med Genet,1986,25 3 : 473
30 Chen D, Xu W, Bales E, et al. SKI activates Wnt/beta- catenin signaling in human melanom. Cancer Res,2003,63 20 : 6 626
31 Aoki K,Aoki M, Sugai M, etal. Chromosomal instability by beta-catenin/TCF transcription in APC or beta-cateninmutant cells. Oncogene,2007,26 24 : 3 511
32 Hanson C A,MillerJ R. Non-traditional roles for the Adenomatous Polyposis Coli(APC)tumor suppressor protein.Gene,2005,361: 1
33田秀芬等,人喉鳞状细胞癌组织中抑癌基因APC蛋白的表达郑州大学学报医学版2008,7(43)716-718
2王正敏,陆书昌,现代耳鼻咽喉科学北京;民军医出版社,2001.10
3 Miwa S, Uchida C, Kitagawa K, et al. Mdm22mediated pRB down regulation is involved in carcinogenesis in a p53-independentmanne. Biochem BiophysResCommun, 2006, 340 1 :54 - 61Weiberg R A The retinoblastoma and cell cycle control 1995(2)
4 Yamasaki L. Role of RB tumor suppressor in cancer. Cance TreatRes,2003, 115( 9) :209 - 239宗永生,王连唐,耳鼻咽喉肿瘤病理部分广州;广东科技出版社,2002.5
5黄选兆等,实用耳鼻咽喉头颈外科学北京;人民卫生出版社,2007.12
6 CAHILLY-SNYDER L,YANG-FENG T,FRANCKE U,et al. moccular analysis and chromosomal mapping of amplified genes isolated from a transformed mouse 3T3 cell line. Somet Cell Genet,1987,13(1):235-244
7 OLINER J D,KINZLER K W,MELTZER P S,et al.Amplification of a gene encoding a P53-associated protein in human sarcomas. Nature,1992,358(2): 80-81
8 Weissman AM. Themes and variations on ubiquitylation. Nat Rev Mol Cell Biol,2001,2 3 :169-178
9 Jin Y, Zeng SX, Dai MS, et al. MDM2 inhibits PCAF (p300/CREB-binding protein-associated factor)–mediated P53 acetylation. J Biol Chem, 2002, 277 (34): 30838-30843
10 Yin Y, Luciani MG, Fahraeus. P53 stability and activity is regulated by Mdm2-mediated induction of alternative p53 translation products. Nat Cell Biol,2002,4 6 : 462-467
11李小华,陈德基,江曼,等.实验性肺癌癌变过程中p53,mdm2,K- ras及PCNA蛋白的表达.癌症, 2001; 20(9) : 921– 925
12任义,吴逸明,董海燕,等.肺癌组织中MDM2,P53蛋白的表达.郑州大学学报医学版,2005; 40(1)67– 69
13孙志,细胞周期分子调控与癌变.国外医学遗传学分册,1996;19(1);50
14张晓辉,细胞周期的网络调控系统Cyclins-CDKs-CKIs及其余肿瘤的关系.国外医学肿瘤学分册,1997;25(1):77
15王斌全等,喉癌组织RB基因突变及其与临床分期的关系临床耳鼻咽喉科杂志2003 ,2 17 :89-90
16 Helt AM,Galloway.Mechannisms by which DNA tumor virus oncoproteins target the Rb family of pocket protein.Carcinogenesis, 2003,24(2);159-169
17 Reichelt M, Zang K D, Seifert M, et al. The yeast two-hybrid system reveals no interaction between p73 alpha and SV40 large T-antigen.Arch Virol,1999,144(3):621-626
18 Junen Gershon T, ShifmanO, UngerT, etal. The MDM2 oncoprotein interacts with the cell fate regulator numb.Mol CellBiol, 1998,18:3974-3982
19 Sdek P, Ying H, Chang D L, et al. MDM2 promote sproteasome-dependent ubiquitin-independent degradation of retinoblastoma protein. MolCell, 2005,9(5):699 - 708 .
1王正敏,陆书昌,现代耳鼻咽喉科学北京:人民军医出版社,2001.10
2周梁,董频,临床耳鼻咽喉头颈肿瘤学上海:复旦大学出版社,2008.9
3姜泗长,耳鼻咽喉-头颈外科手术学北京:人民军医出版社,2005.9
4王正敏,临床耳鼻咽喉科学上海:复旦大学出版社, 2007.10
5宗永生,王连唐,耳鼻咽喉肿瘤病理部分广州;广东科技出版社,2002.5
6黄选兆等,实用耳鼻咽喉头颈外科学北京:人民卫生出版社,2007.12
7王家东,耳鼻咽喉头颈肿瘤外科学上海:上海交通大学出版社,2007.9
8王天铎,喉科手术学北京:人民卫生出版社,2007.4
9杨宝琦,耳鼻咽喉科学新进展天津:天津科学技术出版社,2000.3
10罗荣城,肿瘤生物治疗学北京:人民卫生出版社,2006.08
11桑建中等,P53、Rb和P16在喉鳞癌组织中的表达及意义山东医药,2005,45:17-19
12 Reddel RR. The role of the senescence and immortalizationin carcinogenesis. Carcinogenesis, 2000, 21 3 : 477
13 Cook JR,HillDA, Humphrey PA, et al. Squamous cell car cinoma arising in recurrent respiratory papillomatosis with pulmonary involvement: emerging common pattern of clinical features and human papillomavirus serotype association. Mod Pathol,2000,13 8 : 914
14 Rassekh CH, Rady PL, Arany I, et al. Combined Epstein-Barrvirus and human papillomavirus infection in nasopharyngealcarcinoma. Laryngoscope, 1998,108 3 : 362
15 YAMAUCHI T, WATANABE M,KUBOTA T, et al. Cyclooxygenase22 expression as a new marker for patients with colorectal cancer[J ]. Dis Colon Rectum 2002 ,45:98 - 103
16 TSUJII M , KAWANO S, TSUJI S, et al. Cyclooxy2genase regulates angiogenesis induced by colon cancer cells. Cell ,1998 ,93:705 - 716
17 KAKIUCHI Y, TSUJI S, TSUJII M , et al. Cyclooxygenase-2 activity altered the cell-surface carbohydrate antigens on colon cancer cells and enhanced liver me- tastasis. Cancer Res ,2002 ,62:1567 - 1572
18 YUAN A , YU C J , SHUN C T, et al. Total cyclooxygenase-2 mRNA levels correlate with vascular endothelial growth factor mRNA levels , tumor angiogenesis and prognosis in non-small cell lung cancer patients. Int J Cancer ,2005 ,115:545 - 555
19 HARIZI H, JUZAN M , PITARD V , et al. Cyclooxygenase-2-issued prostaglandin enhances the production of endogenous IL-10 , which down-regulates dendritic cell functions. J Immunol ,2002 ,168:2255 - 2263
20 JEONG M A , LEE K W, YOON D Y, et al. Jaceosidin , a pharmacologically active flavone derived from Artemisia argyi , inhibits phorbol2ester2induced upregulation of COX-2 and MMP-9 by blocking phosphorylation of ERK-1 and-2 in cultured human mammary epithelial cells. Ann N Y Acad Sci ,2007 ,1095:458- 466
21陈望燕等,COX-2基因及产物在人类喉癌组织中的表达
22临床耳鼻咽喉头颈外科杂志,2008.6 , 22:532-535
23李晓光,于肖鹏,赵海兰,等. PCNA在口腔鳞癌中的表达及其临床意义.泰山医学院学报,2005,26 1 : 20 - 22
24李正贤等,PCNA、Bcl-2和Bax在喉癌组织中的表达及其意义临床军医杂志,2008.12,36:850-852
25姚军.p27与卵巢癌.华夏医学,2003,16 2 :125-127
26李学忠等,MDM2与p27在下咽癌中的表达及其临床意义临床耳鼻咽喉头颈外科杂志,2004.12 , 18:716-718
27 DONG J T,LAMB PW,RINKER-SCHAEFFER CW,et al.KAI1,a metastasis suppressor gene for prostate cancer on human chromosome 11p11.2 J.Science,1995,268 (5) :8842886
28白伟良,任重,潘子民,等.KAI1基因在喉鳞癌中的表达及其临床意义.中华肿瘤杂志,2005,27( 5):289-291
29 Herrera L, Kakati S, GibasL, et al. Gardner syndrome in a man with an interstitial deletion of 5q. Am J Med Genet,1986,25 3 : 473
30 Chen D, Xu W, Bales E, et al. SKI activates Wnt/beta- catenin signaling in human melanom. Cancer Res,2003,63 20 : 6 626
31 Aoki K,Aoki M, Sugai M, etal. Chromosomal instability by beta-catenin/TCF transcription in APC or beta-cateninmutant cells. Oncogene,2007,26 24 : 3 511
32 Hanson C A,MillerJ R. Non-traditional roles for the Adenomatous Polyposis Coli(APC)tumor suppressor protein.Gene,2005,361: 1
33田秀芬等,人喉鳞状细胞癌组织中抑癌基因APC蛋白的表达郑州大学学报医学版2008,7(43)716-718