膀胱癌中BLU基因启动子高甲基化及转录水平.doc

膀胱癌中BLU基因启动子高甲基化及转录水平 　 作者：张剑英，李智，于永春，段建敏 【摘要】 目的 研究膀胱癌中BLU基因启动子甲基化状态、转录水平及其意义。方法 采用甲基化特异性PCR(methylation specific PCR, MSP)技术和逆转录PCR(RTPCR)技术分别检测54例膀胱癌患者癌组织及45例相应癌周正常组织、3例非肿瘤患者的正常膀胱组织中BLU基因启动子甲基化状态和转录水平。结果 ①31.5%(17/54) 膀胱癌组织中BLU基因高甲基化，相应癌周正常组织和3例正常膀胱组织均未发现该基因高甲基化改变；②甲基化状态与膀胱癌临床病理参数无明显相关性；③在45例膀胱癌组织中，BLU表达缺失率为42.2%(19/45)；④BLU mRNA表达缺失与肿瘤临床分期和病理分级相关(Plt;0.05)；⑤在启动子高甲基化的膀胱癌中，92.3%(12/13)BLU mRNA表达异常下调或缺失。结论 膀胱癌中频繁发生BLU基因的高甲基化和mRNA表达缺失，其高甲基化可能是基因转录失活的重要原因。 【关键词】 膀胱癌；BLU；甲基化；转录；甲基化特异性PCR 　　Promoter hypermethylation and transcription level of BLU in bladder cancer 　　ABSTRACT: Objective To investigate the promoter methylation status and the transcription level of BLU in bladder carcinoma and their significance. Methods The methylation status of promoter and the transcription level of BLU were examined by methylation specific PCR (MSP) and RTPCR in bladder cancer tissues from 54 patients, adjacent normal tissues from 45 cases and normal bladder tissues from 3 noncancerous patients. Results ① 31.5%(17/54) bladder cancer tissues showed BLU hypermethylation; neither the paired adjacent normal tissues nor the three normal bladder tissues showed BLU hypermethylation. ② There was no significant correlation between the methylation status and the clinical pathological parameters in bladder cancer. ③ The expression of BLU was absent in 42.2% (19/45) of bladder carcinoma from cystectomy. ④ Loss of BLU expression was associated with the stage and grade of the tumor (Plt;0.05). ⑤ The expression of BLU mRNA was abnormally downregulated or absent in 92.3% (12/13) of bladder carcinoma tissues with promoter hypermethylation. Conclusion BLU was found frequently hypermethylated and absent of mRNA expression in bladder carcinoma. Hypermethylation of BLU may contribute to its transcription inactivation. 　　KEY WORDS: bladder cancer; BLU; methylation; transcription; methylation specific PCR (MSP) 　　肿瘤抑制基因启动子区CpG岛的高甲基化已被确立为基因转录失活的一种重要机制[1]。膀胱癌是泌尿系统最常见的恶性肿瘤,近年来的研究发现，包括p16、Ecadherin、VHL、RAS