摘要
目的:应用甲基化特异性PCR(MSP)技术检测胃癌组织中Caveolin-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.
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 protocols. After the treatment with CpGenome DNA Modification Kit, the extracted DNA was amplified using primers either methylated or unmethylated. The PCR products were analyzed by both agarose gel electrophoresis and sequencing. RESULTS： No methylation of the 5＇CpG island in the promoter region of Caveolin-1 gene exon-2 was observed in the 6 samples of normal gastric tissues. In the 30 samples of gastric carcinoma tissues, the methylation positive rate of 5 ＇CpG island in the promoter region of Caveolin-1 gene exon-2 was 90% （27/30） , among which 16 cases （53. 3%） were only amplified by methylated primers and 11 cases （36.7%） were amplified both by methylated and unmethylated primers. The methylation rate of the promoter region of Caveolin-1 gene exon-2 in gastric carcinoma was significantly higher than that in normal gastric tissues. CONCLUSION： The methylation of the 5＇ CpG island in the promoter region of Caveolin-1 gene exists in gastric carcinoma and it may be involved in the early process of gastric carcinogenesis.
出处
《第四军医大学学报》
北大核心
2009年第17期1591-1593,共3页
Journal of the Fourth Military Medical University
基金
全军医药卫生科研基金(06MB102)
甘肃省自然科学基金(3ZS061-A25-094)
