肝癌细胞中OY-TES-1基因启动子的甲基化状态  

Methylation status of CpG sites in the promoter region of the OY-TES-1 gene in hepatoma cell lines

作　　者：王炜炜[1] 范蓉[1] 罗彬[1] 郭文文[1] 张庆梅[1] 林永达[1] 肖绍文[2] 谢小薰[1] 

机构地区：[1]广西医科大学组织胚胎学教研室广西区域高发肿瘤防治研究重点实验室,广西壮族自治区南宁市530021 [2]广西医科大学第一临床医院神经外科,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2011年第21期2274-2278,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30760055;No.81060207;广西教育厅创新团队基金资助项目;No.BGXZ2007010;广西自然科学基金资助项目;No.0832144;No.2011GXNSFA018275;广西研究生科研创新基金资助项目;No.2010105981001M228~~

摘　　要：目的:研究肝癌细胞株OY-TES-1启动子的甲基化状态及甲基转移酶抑制剂对其启动子甲基化的影响.方法:运用生物信息学在线软件预测OYTES-1启动子区域及转录因子结合位点;应用亚硫酸氢盐测序法(bisulfite-sequencingPCRBSP)分析目的基因CpG位点的甲基化状态用甲基转移酶抑制剂5-Aza-CdR干预肝癌细胞株BEL-7404,比较干预前后OY-TES-1启动子甲基化状态的改变.结果:通过软件分析确定了OY-TES-1启动子区域位于-184-+67bp,该区域含多个潜在的转录因子结合位点;在6种肝癌细胞株中OY-TES-1启动子的甲基化频率范围为51.25%-87.92%;肝癌细胞株BEL-7404经过5-Aza-CdR干预后,甲基化频率由干预前的87.08%下降至21.25%.结论:在检测的肝癌细胞株中,OY-TES-1启动子呈高甲基化状态,甲基转移酶抑制剂能显著降低肝癌细胞株OY-TES-1启动子甲基化频率.AIM： To detect the methylation status of CpGsites in the promoter region of the OY-TES-1 gene in six hepatorna cell lines and to investigate the influence of DNA methyltransferase inhibi- tor 5-Aza-CdR on methylation status of OY- TES-1 promoter in hepatoma cell line BEL-7404. METHODS： Online bioinformatic tools were used to predict the promoter region of the OY- TES-1 gene and putative transcription factor binding sites. Bisulfite sequencing PCR （BSP） was applied to detect the methylation status of CpG sites in the promoter region of the OY- TES-1 gene. After treatment of BEL-7404 cells with 5-Aza-CdR, the methylation status of CpG sites in the promoter region of OY-TES-1 gene was detected again. RESULTS： The promoter region of the OY- TES-1 gene was predicted to be located between -184 bp and ＋67 bp, which contains several potential transcription factor binding sites. The frequency of promoter methylation of the OY- TES-1 gene ranged between 51.25% and 87.92% in six hepatoma cell lines. 5-Aza-CdR treatment decreased the frequency of promoter methyla- tion of the OY-TES-1 gene from 87.08% to 21.25% in BEL-7404 cells. CONCLUSION： The OY-TES-1 gene shows rela- tively high frequency of promoter methylation in six hepatoma cell lines. DNA methyltrans- ferase inhibitor could significantly decrease the frequency of promoter methylation of the OY- TES-1 gene in hepatoma cell line BEL-7404.

关 键 词：肝癌 启动子 甲基化 OY—TES-1 

分 类 号：R735.7[医药卫生—肿瘤]