胃癌组织中Caveolin-1基因外显子2甲基化状态检测

目的：应用甲基化特异性PCR（MSP）技术检测胃癌组织中Caveotin-1基因外显子2启动子区域5’端CpG岛甲基化状况，探讨Caveolin-1基因甲基化在胃癌发生发展中的作用．方法：收集30例胃癌组织和6例距肿瘤5cm以上的癌旁正常胃组织，运用标准蛋白酶K消化、酚-氯仿抽提法提取组织中基因组DNA，采用CpGenome DNA Modification Kit对DNA进行修饰后以甲基化特异性引物及非甲基化引物进行PCR，通过琼脂糖凝胶电泳及产物测序判定结果．结果：6例癌旁正常胃组织Caveolin-1基因外显子2启动子区域5＇端CpG岛均为甲基化阴性．30例胃癌组织中有27例甲基化阳性，甲基化率为90％（27／30）．其中16例胃癌组织（53．3％）仅有甲基化引物扩增出目的条带，表现为完全甲基化；11例胃癌组织（36．7％）甲基化与非甲基化引物均扩增出目的条带，表现为部分甲基化．统计结果显示胃癌组织中Caveolin-1基因外显子2启动子区域甲基化率显著高于癌旁正常胃组织．结论：胃癌组织中存在Caveolin-1基因外显子2启动子区域高甲基化，且可能参与胃癌的早期过程．

Detection of methylation of Caveolin-1 gene exon 2 in gastric carcinoma

AIM:To detect methylation of 5'CpG island in the promoter region of Caveolin-1 gene exon-2 in gastric carcinoma by methylation-specific PCR(MSP) technique and to analyze the role of methylation of Caveolin-1 gene in the genesis and development of gastric carcinoma.METHODS:Thirty samples of gastric carcinoma tissues and 6 samples of normal gastric tissues taken at least 5 cm away from tumors were collected.DNA of the tissues was extracted by standard proteinase K digestion and phenol/chloroform extraction pr...