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）.
出处
《解剖学杂志》
CAS
CSCD
北大核心
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）
