ANXA7基因异常甲基化与神经胶质瘤发生相关性的研究

Significance of ANXA7 gene promoter methylation detection in glioblastoma

目的研究ANXA7基因异常甲基化与神经胶质瘤发生的相关性。方法采用硫化测序PCR(BS-PCR)引物以及DNA甲基化特异性PCR(MS-PCR)引物检测ANXA7基因在人脑神经胶质组织及正常脑组织中的表达差异,随后经PCR扩增进行电泳以及测序分析。结果 5例健康标本及5例神经胶质瘤患者标本DNA经硫化且BS-PCR测序分析,其健康标本甲基化率分别为1.5%、1%、1%、1.5%、1%,神经胶质瘤患者甲基化率分别为92%、78.5%、86%、56%、90%;MS-PCR分析结果显示:ANXA7基因在24例正常脑组织中呈完全非甲基化状态,在60例神经胶质瘤患者中其甲基化阳性率为46.67%(p=0.000);高级别神经胶质瘤患者甲基化阳性率(56.67%)高于低级别组(26.67%)(P=0.018)。结论 ANXA7基因异常甲基化可能参与神经胶质瘤的发生发展。

Objective The aim of this study was to evaluate the significance of ANXA7 gene promoter hypermethylation in Glioblastoma. Methods Sixty patients with Glioblastoma and 24 healthy donors were adopted in this study. DNA was isolated from cerebra tissue, and BS-PCR and MS-PCR methods were used to detect the status of ANXA7 gene methylation in health donors and newly diagnosed Glioblastoma patients in using PCR amplication and sequencing. Results The results indicated that the methylated rate of the ANXA7 gene in 5 healthy donors were 1.5%, 1%, 1%, 1.5%, 1% by BS-PCR and sequencing analysis, however, the methylated rate of the ANXA7 gene in 5 Glioblastoma patients were 92% ,78.5% ,86% ,56% ,90%. MS-PCR analysis showed the ANXA7 gene was unmethylated in cerebra tissue samples from health donors. Among 60 newly diagnosed Glioblastoma patients, 25 patients were found in ANXA7 gene methylation by MS-PCR, the positive rate was 41.67% （p = 0.000）. Furthermore, the positive rate of ANXA7 methylation in the group of high stage disease was higher than in the group of low stage. Conclusions The aberrant methylation of the ANXA7 gene was perhaps involved in the occurrence of glioblastoma.