胶质瘤死亡相关蛋白激酶1基因甲基化检测及临床意义  

Detection of Dapk1 gene promoter methylation and its clinical value in gliomas

作　　者：王涛 梁日初 刘小飞 何健 廖泉 周佳[2] Wang Tao;Liang Richu;Liu Xiaofei;He Jian;Liao Quan;Zhou Jia(Department of Neurosurgery,the Second Affiliated Hospital of South China University,Hengyang 421001,China;Department of Emergency,the First Affiliated Hospital of South China University,Hengyang 421001,China)

机构地区：[1]南华大学附属第二医院神经外科,衡阳421001 [2]南华大学附属第一医院急诊科,衡阳421001

出　　处：《中华实验外科杂志》2020年第5期933-936,共4页Chinese Journal of Experimental Surgery

基　　金：湖南省卫生健康委重点指导项目(20201912);衡阳市科技局2019年指导性项目(132)。

摘　　要：目的:检测胶质瘤中死亡相关蛋白激酶1基因(Dapk1)甲基化并探讨其临床意义。方法:分别应用甲基特异性聚合酶链反应(PCR)、反转录-聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blot)检测2015年1月至2017年1月南华大学附属第一医院和南华大学附属第二医院诊治的70例胶质瘤患者(GLA组)、40例颅脑良性疾病患者(CON组)、U251和BT325胶质瘤细胞(购于中国科学院细胞库)Dapk1基因甲基化、mRNA和蛋白质比较,采用χ2检验比较胶质瘤患者不同临床病理中Dapk1基因甲基化的差异,Kaplan-Meier法对胶质瘤患者Dapk1基因甲基化和未甲基化患者生存分析。结果:GLA组胶质瘤患者Dapk1基因甲基化率显著高于CON组颅脑良性疾病患者(65.7%比2.5%, χ2=39.023, P<0.05)。U251和BT325胶质瘤细胞中Dapk1基因处于甲基化状态。GLA组胶质瘤患者、U251和BT325胶质瘤细胞中Dapk1基因mRNA和蛋白表达均显著低于CON组颅脑良性疾病患者(mRNA相对表达量分别为0.34±0.05、0.35±0.06、0.33±0.05、0.65±0.08,蛋白质相对表达量分别为0.22±0.04、0.23±0.05、0.21±0.04、0.57±0.07, F=67.125、52.198, P<0.01)。GLA组胶质瘤患者Dapk1基因甲基化率在肿瘤最大径和世界卫生组织(WHO)分级方面的差异均有统计学意义(甲基化率分别为78.9%、80.0%, χ2=5.240、7.039, P值均<0.05),肿瘤最大径≥5 cm和WHO Ⅲ+Ⅳ级患者Dapk1基因甲基化率显著高于肿瘤最大径<5 cm和WHO Ⅰ+Ⅱ级级患者。Dapk1基因甲基化患者中位生存期显著低于未甲基化患者[(13.8±1.1)个月比(16.3±1.5)个月,Logrank=5.107, P<0.05]。 结论:Dapk1基因高甲基化与胶质瘤发病机制有关,有助于胶质瘤病情及预后评估。Objective To detect the death-associated protein kinase 1(Dapk1)gene methylation and study its clinical value in gliomas.Methods The methylation status of Dapk1 gene,mRNA and protein expression in gloma group(70 patients with gliomas),CON group(40 patients with benign brain disease),U251 and BT325 glioma cells was detected by methyl-specific polymerase chain reaction(PCR),reverse transcriptase-PCR(RT-PCR)and Western blotting.The relationship between clinicopathological factors and Dapk1 gene methylation was analyzed.The survival of glioma patients with or without Dapk1 gene methylation was studied and compared by Kaplan-Meier survival analysis.Results The methylation rate of Dapk1 gene in glioma group was significantly higher than in CON group(65.7%vs.2.5%,χ2=39.023,P<0.05).The Dapk1 gene was methylated in U251 and BT325 glioma cells.The expression of Dapk1 gene mRNA and protein in glioma group,U251 and BT325 cells was significantly lower than that in CON group(mRNA=0.34±0.05,0.35±0.06,0.33±0.05,0.65±0.08,protein=0.22±0.04,0.23±0.05,0.21±0.04,0.57±0.07,F=67.125,52.198,P<0.01).The Dapk1 gene methylation rate in glioma patients was significantly correlated with maximum tumor diameters and World Health Organization(WHO)staging(methylation rate=78.9%,80.0%,χ2=5.240,7.039,P<0.05).The median survival time in glioma patients with Dapk1 gene methylation was significantly shorter than in patients with Dapk1 gene unmethylation[(13.8±1.1)m vs.(16.3±1.5)m,Logrank=5.107,P<0.05].Conclusion The hypermethylation of Dapk1 gene is related to the pathogenesis of glioma,which can provide evidence of gene level for the evaluation of glioma disease and prognosis.

关 键 词：胶质瘤 死亡相关蛋白激酶1基因 甲基化 临床意义 

分 类 号：R739.41[医药卫生—肿瘤]