全反式维A酸对卵巢上皮性腺癌细胞株COC2中RARa基因甲基化影响的研究  

Influence of all-trans retinoic acid on the methylation of the RARa gene in ovarian cancer cell line COC2

作　　者：居艳[1] 刘琦[1] 吴元赭[1] 林静娴[1] 

机构地区：[1]南京军区南京总医院妇产科,江苏南京210002

出　　处：《医学研究生学报》2009年第5期472-475,479,I0007,共6页Journal of Medical Postgraduates

基　　金：南京军区医药卫生科研基金资助项目(批准号:07M084)

摘　　要：目的:全反式维A酸是一种经典的诱导肿瘤细胞分化的药物,但关于其对肿瘤细胞的异常甲基化的影响,国内研究仍很少,文中通过观察全反式维A酸(ATRA)对卵巢上皮性腺癌细胞株COC2中RARa基因启动子甲基化位点的影响,及其对RARa基因mRNA表达的影响,探讨ATRA对RARa基因启动子区异常甲基化可能的作用。方法:用不同浓度及作用时间的ATRA体外干预COC2细胞,甲基化特异性PCR及基因测序法检测用药前后COC2细胞中RARa启动子区甲基化状态的改变。RT-PCR法检测各组RARa基因mRNA表达的改变。结果:COC2细胞中存在RARa基因启动子区异常甲基化;不同浓度ATRA处理后,RARa启动子的异常甲基化位点出现部分逆转,并在一定范围内表现出浓度及时间依赖性,RARa基因的mRNA表达也同样表现为增高趋势。结论:COC2细胞中存在RARa基因启动子区异常高甲基化;ATRA可以部分逆转RARa启动子区的异常甲基化位点并增加RARamRNA的表达。Objective ： All-trans retinoic acid （ATRA） is a classic drug that can induce tumor differentiation, and its influence on the aberrant methylation of cancer cells has been studied insufficiently. The objective of this study was to observe the influence of ATRA on the methylation of the RARa gene promoter in ovarian cancer cell line COC2 and its relationship with the RARa expression. Methods ： The ovarian cancer cell line COC2 was treated with different concentrations of ATRA for different times. Methyla- tion specific PCR （MSP） and bisulfate sequencing methods were used to detect the changes in the methylation of the RARa promoter after ATRA treatment. RT-PCR was employed to observe the changes in the expression level of RARa mRNA. Results ： Aberrant methylation of the RARa gene promoter was found in the ovarian cancer cell line COC2. ATRA treatment decreased the number of RARa promoter methyla-tion sites in COC2 within a certain scope in a concentration- and time-dependent manner, and increased the expression of RARa mRNA. Conclusion ： Aberrantly high methylation of the RARa promoter exists in the ovarian cancer cell line COC2; ATRA can partially reverse the methylation and increase the RARa mRNA expression.

关 键 词：全反式维A酸 卵巢上皮性腺癌细胞 RARa基因 甲基化 

分 类 号：R737.31[医药卫生—肿瘤]