应用焦磷酸测序法检测肝癌中4种基因甲基化的研究  

Pyrosequencing detection Cp G methylation of 4 genes in heptocarcinoma patients

作　　者：牟瑛[1] 宋添添[1] 宫雪[1] 李宏伟[2] 欧阳晶 

机构地区：[1]解放军第三〇二医院营养科,北京100032 [2]第四军医大学药学系药物基因组学教研室全军基因芯片重点实验室,陕西西安710033 [3]海军总医院医务部医疗科,北京100048

出　　处：《中国热带医学》2015年第6期664-667,共4页China Tropical Medicine

摘　　要：目的使用焦磷酸测序技术检测GSTP1、E-cadherin、RARβ、P16基因启动子区域在肝癌组织中的甲基化状态,为临床早期诊断肝癌奠定基础。方法取肝癌组织及肝硬化组织石蜡切片标本,使用焦磷酸测序仪检测GSTP1、E-cadherin、RARβ、P16基因启动子甲基化状态。结果肝癌与肝硬化组织相比较GSTP1基因超甲基化率为20%(4/20),E-cadherin基因超甲基化率为40%(8/20),RARβ基因超甲基化率为55%(11/20),P16基因超甲基化率为75%(15/20)。其中E-cadherin、RARβ、P16基因有统计学差异(P<0.05)。结论 E-cadherin、RARβ、P16基因在肝细胞癌患者血清中的异常甲基化表达程度,可作为肝癌早期辅助诊断的分子标志物,焦磷酸测序技术检测技术为实现临床早期诊断、早期肝癌治疗奠定了试验基础。Objective To establish and evaluate a quantitative method to analyze genomic NDA methylation level for early clinical diagnosis of hepatocellular carcinoma and discover the methylative level difference between hepatocellular carcinoma and cirrhosis tissues. Methods The tissue specimens were obtained from patients with hepatocellular carcinoma and cirrhosis and the methylation status of GSTP1, E-cadherin, RARI3, P16 gene promoter regionssing were determined by pyrosequencing technology. Results Compared with cirrhosis tissues, the hypermethylative rate of GSP1, E-cadherin, RAR- 13, P16 genes were 20%, 40%, 55% and 75% in hepatocellular carcinoma patients respectively. The hypermethylative rates of E-eadherin, RAR-13, P16 genes showed significant statistical difference compared to that of cirrhosis （P〈0.05）. Conclusion The methylation levels of E-cadherin, RAR-13, P16 genes in hepatocellular carcinoma can be recognized as an useful markers for early diagnosis of hepatocellular carcinoma. Pyrosequencing technology is an quick, sensitive and high throughput detection method.

关 键 词：焦磷酸测序 启动子甲基化 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]