OPCML、DAPK基因甲基化与宫颈癌CasKi细胞凋亡相关性的研究  

Study on the relevancy between OPCML and DAPK gene methylation and CasKi cell apoptosis

作　　者：王艳华[1] 黄利鸣[2] 

机构地区：[1]三峡大学医学院生物研究所,湖北宜昌443002 [2]三峡大学医学院病理学教研室,湖北宜昌443002

出　　处：《中国药理学通报》2008年第4期543-546,共4页Chinese Pharmacological Bulletin

基　　金：湖北省科技厅科学研究计划项目(No2005AA40IC57)

摘　　要：目的研究天花粉蛋白(TCS)诱导宫颈癌CasKi细胞凋亡过程中肿瘤抑制基因OPCML、DAPK甲基化的变化情况,并探讨其在宫颈癌的发生发展中的作用及细胞凋亡和抑癌基因甲基化的相关性,寻找新型的去甲基化药物。方法①应用MTT法检测TCS对CasKi细胞增殖抑制的影响;流式细胞仪分析天花粉蛋白对宫颈癌CasKi细胞诱导凋亡作用;②应用甲基化特异性PCR(MSP)技术检测人宫颈癌细胞OPCML和DAPK基因启动子CpG岛甲基化的状况及天花粉蛋白的去甲基化作用。结果TCS对宫颈癌CasKi细胞生长有明显抑制作用,流式细胞仪检测见特征性的亚二倍体凋亡峰;MSP检测表明宫颈癌CasKi细胞OPCML、DAPK基因启动子区CpG岛呈高度的甲基化状态,TCS处理后OPCML、DAPK基因启动子区CpG岛无异常甲基化表现。结论天花粉蛋白具有抑制宫颈癌CasKi细胞生长、诱导Cas-Ki细胞凋亡和对CasKi细胞OPCML、DAPK基因明显的去甲基化作用;其抑制生长和诱导凋亡作用可能与对OPCML、DAPK基因的去甲基化作用有关。TCS有可能是一种新型的甲基化抑制剂。Aim To study tumor suppressor gene OPC- ML, DAPK methylation changes during the process of CasKi cell apoptosis induced by trichosanthin, and to explore the correlation and the role between cervical cancer cell apoptosis and tumor suppressor gene methylation so as to find new demethylation drugs. Methods ①MTT was applied to assay the inhibition of TCS to CasKi cell proliferation and flow cytometry was used to analyze cervical CasKi cell apoptosis induced by trichosanthin ; ② Methylation-specific PCR（MSP） technology was applied to detect cervical cancer and during cell apoptosis process OPCML and DAPK gene promoter methylation status of CpG islands. Results In CasKi cervical cancer cells, OPCML and DAPK gene promoter region showed a high degree of CpG island methylation status, by trichosanthin treatment, the growth of CasKi markedly was inhibited, and flow cytometry analysed the characteristic sub-G1 peak, OPCML and DAPK gene promoter region showed no CpG island methylation of performance. Conclusions During the process of CasKi cell apoptosis induced by trichosanthin, OPCML and DAPK gene demethylated significantly, accordingly, trichosanthin might be a new methylation inhibitor, and there might be some correlation between cell apoptosis and tumor suppressor gene methylation. And OPCML and DAPK gene methylation tests might become new clinical indicators for the early detection of cervical cancer.

关 键 词：OPCML基因 DAPK基因 天花粉蛋白 CPG岛 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]