膀胱癌细胞凋亡与MDM2和P53基因表达的意义

膀胱癌细胞凋亡与MDM2和P53基因表达的意义 作者：杨承纲，张丽娟，李梅，陈芸 【关键词】 膀胱肿瘤；P53基因；MDM2基因；免疫组织化学；细胞凋亡 ［摘 要］ 目的：研究膀胱癌细胞凋亡、MDM2、P53基因表达的意义。方法：采用免疫组织化学方法及TUNEL法检测11例“正常膀胱黏膜”和83例膀胱移行细胞癌中MDM2、P53基因的表达及细胞凋亡指数。结果：P53阳性率为50.6%,MDM2阳性率为59.03%，AI=1.433 5±0.386 3。MDM2阳性表达与病理分级及临床分期呈负相关(R=-0.263,P=0.016及R=-0.388,P=0.001);P53阳性表达与病理分级及临床分期呈正相关(R=0.492,P=0.001；R=0.341，P＜0.01);MDM2、P53三种蛋白间表达无相关性；AI与MDM2呈负相关（R=-0.226,P=0.042）；AI与病理分级及临床分期呈正相关(R=0.642,P=0.000；R=0.455，P=0.000)；AI与P53无相关性。AI、P53、MDM2的表达与5 a生存率有关。结论：联合运用临床分期及病理分级并检测P53、MDM2可用于判断膀胱移行细胞癌的预后。 　　［关键词］ 膀胱肿瘤；P53基因；MDM2基因；免疫组织化学；细胞凋亡 　　Clinical Significance of Expression of MDM2 P53 Protein and Apoptosis in Bladdercancer 　　Abstract：Objective To investigate the expression of MDM2 Pgp protein and apoptosis and its significance in bladder cancers.Methods Apoptosis Index(AI) was assessed with TUNEL technique,and immunohistochemical stainings of MDM2 and Pgp were performed in 83 cases of the carcinomas and 11 “normal” bladder mncosa.Results The positive expression rate of P53 gene was 50.6% and MDM2 was 59.03%,and AI=1.433 5±0.386 3.The level of MDM2 was negative correlated with the tumor pathological grade and clinical stage(R=-0.263,P=0.016及R=-0.388,P=0.001);The level of P53 was positive correlated with the tumor pathological grade(R=0.492,P=0.001);No correlated was found between MDM2 and P53 gene;The level of AI was negative correlated with MDM2(R=-0.226,P=0.042);The level of AI was positive correlated with the tumor pathological grade and clinical stage (R=0.642,P=0.000；R=0.455，P=0.000);The level of AI was no correlated with P53.AI and the expression of P53 and MDM2 were related to 5year survival rate.Conclusion By exerting the tumor pathological grade and clinical stage and detecting P53 MDM2 genes expressions,the prognosis of bladder cancer can be decided.论文网在线 　　Key words：Bladder neoplasm；P53 gene；MDM2 gene；Immunohistochemistry；Apoptosis MDM2基因是一原癌基因，与P53构成降解反式激活循环通路，参与细胞生长抑制、凋亡、细胞周期调控等过程。细胞凋亡是由基因调控的细胞主动死亡过程，同肿瘤的发生发展以及临床治疗有密切的关系。本研究采用免疫组化方法观测MDM2、P53基因在人膀胱癌细胞中的表达及用TUNEL法检测细胞凋亡，探讨它们在膀胱癌的发生发展中的作用及其对膀胱癌生物学行为和预后的影响。 　　1 材料与方法 　　1.1 材料 收集本院病理科1996年至2004年间膀胱移行细胞癌83例及11例正常膀胱黏膜组织，83例膀胱移行细胞癌中男性66例，女性17例，年龄35岁~82岁，平均年龄58.5岁。病理检查前均未接受过放疗和化疗，术后均进行化疗药物膀胱灌注。按1999年WHO病理组织学分级标准，Ⅰ级29例，Ⅱ级31例，Ⅲ级23例；按UICC临床分期，TisT1期61例，T2T4期22例。所有标本均经10%甲醛固定、石蜡包埋，4 μm厚连续切片。术后随访16个月~5 a，平均38个月，失访5例，肿瘤复发或转移56例，死亡2例，无复发或转移20例。 　　1.2 方法 　　1.2.1 阴性、阳性检测 MDM2、P53单抗均购自福州迈新公司。采用SP法进性免疫组化染色，严格按照试剂盒说明书操作。用已知阳性切片作阳性对照，用PBS代替一抗作阴性对照。结果判定：MDM2、P53以细胞核中出现棕黄色颗粒为阳性细胞，阳性细胞＞10%为阳性结果，反之为阴性结果（见图1，图2）。