RASSF1A基因在膀胱移行细胞癌中转录表达和启动子甲基化研究.docVIP

RASSF1A基因在膀胱移行细胞癌中转录表达和启动子甲基化研究 　　用半定量逆转录聚合酶链反应(RT-PCR)技术鉴测37例膀胱移行细胞癌(bladder transitional cell Carcinoma,BTCC)组织和8例正常膀胱组织中RASSF1A基因mRNA表达水平;用甲基化特异性PCR(Methylation-Specific PCR,MSP)。方法 研究膀胱移行细胞癌组织和正常膀胱组织中的RASSF1A基因启动子区CPG岛甲基化状态。结果 RASSF1AmRNA在8份正常膀胱组织中全部表达,在BTCC组织中的表达率为35.1%(13/37),二组间表达差异有显著性(P0.05)。在正常膀胱组织中RASSF1A基因启动子未发生甲基化,但在BTCC组织中RASSF1A基因启动子甲基化频率为51.3%(19/37)。二组间表达差异有显著性(P 　　【关键词】膀胱移行细胞癌;DNA甲基化;聚合酶链反应;RASSF1A基因 　　【Abstract】Objective To investigate the expression of RASSF1A mRNA and promoter hypermethylation of RASSF1A gene in bladder transitional cell Carcinoma and to analyze the association between them. Methods RASSF1A mRNA expression level was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) in 8 normal bladder tissues and 37 BTCC tissues. The promoter methylation status of RASSF1A gene in BTCC and normal bladder tissues were tested by MSP. Results RASSF1A mRNA were all shown in 8 normal bladder tissues and the expression rate is 35.1% (13/37) in BTCC tissues. There was significantly different between the two groups (P0.05). The frequence of promoter methylation of RASSF1A gene was 51.3% (19/37) in BTCC tissues. But it was not found that the promoter methylation in normal bladder tissues. Amoung the 19 patients with the aberrant methylation,16 showed the loss .expression of RASSF1A mRNA . Conclusion Promoter aberrant hypermethylation may Contribute to the loss or low level of RASSF1A mRNA expression and the genesis of bladder tumor. However,it may be has no correlation with the invasiveness of tumor. 　　【Key words】Bladder transitional cell carcinoma; DNA methylation; Polymerase chain reaction; RAS association domain family 1A gene 　　 　　 　　Ras相关区域家族1A(Ras association domain family1A gene,RASSF1A)是2000年从3号染色体短臂上克隆出来的新型肿瘤抑制基因,位于人类染色体3p21.3位点[1],其抑癌机制研究仍处于初级阶段,且国内报道甚少。本研究采用未定量逆转录聚合酶链反应(RT-PCR)方法研究RASSF1AmRNA在膀胱细胞癌组织、正常膀???组织中的表达,同时观察该基因启动子甲基化状态,分析基因表达与启动子甲基化及与肿瘤发生、发展的关系。 　　1 材料与方法 　　1.1 一般资料收集郑州大学一附院及中牟县人民医院泌尿外科200