卵巢癌细胞中OY-TES-1基因启动子CpG位点甲基化状态分析  

CpG site methylation analysis of OY-TES-1 promoter in ovarian cancer cell ~

作　　者：范蓉[1] 黄巍[2] 郭文文[3] 王炜炜[3] 罗彬[3] 宋慧[3] 肖绍文[3] 谢小薰[3] 

机构地区：[1]广西中医学院组织学与胚胎学教研室,南宁530001 [2]广西中医学院第一附属院妇科,南宁530023 [3]广西医科大学组织学与胚胎学教研室,南宁530021

出　　处：《解剖学杂志》2011年第6期763-765,777,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金（30760055）;广西高发疾病研究创新团队基金（桂教人E2007-591号）;广西中医学院课题（zD200901;P2009079）

摘　　要：目的：了解卵巢癌SKOV3细胞中OY-TES-1启动子区域CpG位点的甲基化状态。方法：运用免疫细胞化学法检测SKOV3细胞中OY-TES-1蛋白表达；通过硫化测序PCR法，分析SKOV3细胞中OY-TES-1启动子区域CpG位点的甲基化状态，并与睾丸组织相比较。结果：免疫细胞化学法检测结果显示，SKOV3细胞中OY-TES-1蛋白呈阳性反应；在检测的OY-TES-1启动子区域（转录起始点-127bp-＋110bp，含24个CpG位点）中，CpG位点的甲基化频率为80．21％，其甲基化状态明显高于睾丸组织。结论：SKOV3细胞中OY-TES-1启动子CpG位点处于高甲基化状态，进一步可探讨甲基化抑制剂氮杂脱氧胞嘧啶对OY-TES-1甲基化状态及其表达水平的影响。Objective： To investigate CpG site methylation of OY-TES-1 promoter in ovarian cancer cell line SKOV3. Methods： OY-TES-1 expression in SKOV3 was detected by immunocytochemical staining with anti-OY-TES-1 antibody. The DNA fragment （-127 bp-＋ll0 bp, including 24 CpG sites） of 5＇ flanking of OY-TES-1 gene was amplified from the ge- nomic DNA extracted from SKOV3 by bisulfite sequencing PCR （BSP）. CpG site methylation of OY-TES-1 promoter was compared with testis. Results： Positive expression of OY-TES-1 protein was found in SKOV3. Methylation percentage of 24 CpG sites on OY-TES-1 promoter tested was 80.21% in SKOV3, which was higher than that in normal testis tissues. Conclusion： OY-TES-1 has hypermethylated promoter CpG sites in SKOV3. Further study needs to learn the effect of these CpG sites methylation on the expression of OY-TES-1 when using DNA methyltransferase inhibitor 5-aza-2Cdeoxycytidine （DAC）.

关 键 词：癌-睾丸抗原 卵巢癌 CPG 甲基化 OY-TES-1 

分 类 号：R394[医药卫生—医学遗传学]