Claudin1基因在大肠癌中表达检测意义.docVIP

Claudin1基因在大肠癌中表达检测意义 　　[摘要] 目的 探讨Claudin-1基因mRNA在大肠癌发病过程的表达及意义。方法 收集62例液氮保存的新鲜大肠癌组织及其对应的正常大肠组织,常规方法提取组织总RNA。将这些总RNA逆转录成cDNA后,利用实时荧光定量PCR方法检测Claudin-1基因在这些组织中的表达情况,最后统计分析Claudin-1表达变化与大肠癌进展相关的临床病理参数之间的关系。结果 与正常大肠组织相比,Claudin-1基因在大肠癌组织中表达明显增强,此外,DukesⅢ、Ⅳ期大肠癌患者Claudin-1基因表达比Ⅰ、Ⅱ期患者明显增强。结论 Claudin-1基因在大肠癌中高表达,提示其可能与大肠癌发生和进展过程有关。 　　[关键词] 大肠癌;Claudin-1;实时荧光定量PCR 　　[中图分类号] R735.3+4[文献标识码] A[文章编号] 1673-9701(2010)14-15-03 　　 　　Detection of Claudin-1 mRNA Expression in Large Intestine Cancer 　　HU Jingping 　　Gastroenterology,Yongzhou City Peoples Hospital,Yongzhou 425000,China 　　 　　[Abstract] Objective To explore the expression change of Claudin-1 mRNA and its significance in the pathogenesis of large intestine cancer. Methods Total RNA of 62 fresh large intestine cancer samples and their corresponding normal samples was extracted by routine Trizol way. After total RNA reverse transcription into cDNA,the Claudin-1 gene expression in these samples was detected by real-time PCR. Finally,the correlation of Claudin-1 expression with clinical patho-parameters was analyzed in large intestine cancer. Results Compared with the normal samples,the Claudin-1 expression was markedly increased in large intestine cancer samples. In addition,the Claudin-1 expression in large intestine cancer samples of Dukes Ⅲ and Ⅳ phases was obvious higher than that in Dukes ⅠandⅡ phases. Conclusion Over expressed Claudin-1 may promote the pathogenesis of large intestine cancer. 　　[Key words] Large intestine cancer;Claudin-1;Real-time PCR 　　大肠癌是目前包括中国在内国际上最常见的恶性肿瘤,其发生、发展受到遗传、环境以及饮食结构等众多因素影响,出现一些重要基因表达变异,最终导致肿瘤发生。 　　Claudin蛋白家族包括24个成员,其表达具有组织特异性,在维持细胞极性和紧密连接屏障功能方面起着重要作用。一些研究表明,Claudin蛋白表达异常与一些恶性肿瘤的发生和发展及预后密切相关。Claudin1蛋白是1998年Furuse等[1]从鸡肝中得到了一种紧密连接的蛋白质片段,其相对分子质量为22Kd。目前已有少数研究表明,Claudin-1变异的蛋白参与了肿瘤的发病过程,其中也包括大肠癌。但这种研究基本都是基于免疫组化的方法来验证的,很少在mRNA水平上来证实。因此本研究主要利用实时荧光定量PCR的方法来探讨Claudin-1基因变异的mRNA表达在大肠癌发病过程中的意义,并报道如下。 　　1材料与方法 　　1.1材料 　　1.1.1主要试剂逆转录试剂盒及实时荧光定量PCR试剂盒购自北京天根公司。Trizol从日本 Takara公司购买。