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目的了解胃癌中p16、E-cadherin、Runx3、DAPK和CHFR基因启动子CpG岛的甲基化状态,并探讨基因异常甲基化与mRNA表达的关系及其意义。方法采用甲基化特异性PCR(MSP)技术检测12例胃癌患者癌组织及其相应的癌旁正常组织中5种基因的甲基化状态,以12例胃镜活检的正常胃组织作为对照,采用RT-PCR方法相应检测了5种基因的mRNA表达水平。并分析每种基因甲基化程度与表达水平之间的关系。结果胃癌组织中,p16、E-cadherin、Runx3、DAPK和CHFR基因的甲基化率分别为41.7%、8.3%、58.3%、33.3%和58.3%,相应的癌旁正常组织中,p16、E-cadherin、Runx3、DAPK和CHFR基因的甲基化率分别为8.3%、0.0%、8.3%、8.3%和16.7%,且75%的胃癌组织中至少有一种基因甲基化,显著高于癌旁正常组织25%(P<0.05),而健康对照组织中无基因甲基化,在甲基化的胃癌组织中均无p16、E-cadherin和DAPKmRNA表达,而癌旁正常组织、非甲基化的癌组织中均有其表达。结论胃癌中多基因启动子CpG岛高甲基化是一个频繁的事件,并且基因启动子高甲基化与其mRNA表达缺失有关,这可能参与了胃癌的发生发展。
Objective To investigate the methylation status of CpG islands in promoters of p16, E-cadherin, Runx3, DAPK and CHFR in gastric cancer and to explore the relationship between abnormal methylation and mRNA expression. Methods Methylation-specific PCR (MSP) was used to detect the methylation status of five genes in 12 cases of gastric cancer and their adjacent normal tissues. 12 cases of normal gastric tissue biopsies were used as control. The mRNA expression levels of five genes were detected by RT-PCR. And analyze the relationship between the methylation level and the expression level of each gene. Results The methylation rates of p16, E-cadherin, Runx3, DAPK and CHFR in gastric cancer tissues were 41.7%, 8.3%, 58.3%, 33.3% and 58.3%, respectively. The methylation rates of -cadherin, Runx3, DAPK and CHFR genes were 8.3%, 0.0%, 8.3%, 8.3% and 16.7% respectively, and methylation of at least one of 75% of gastric cancer tissues was significantly higher than that of cancer Normal tissue was 25% (P <0.05), while no gene methylation was found in healthy control tissues. There was no expression of p16, E-cadherin and DAPK mRNA in normal gastric tissue, The cancerous tissue has its expression. Conclusion The hypermethylation of multi-gene promoter CpG island in gastric cancer is a frequent event, and the hypermethylation of gene promoter is related to the loss of its mRNA expression, which may be involved in the occurrence and development of gastric cancer.