CHFR在子宫内膜癌细胞中的表达和甲基化调控  

Expression and methylation regulation of CHFR in endometrial cancer cells

作　　者：王小韵[1] 杨越波[1] 李小毛[1] 许成芳[1] 方莉[1] 张旭[1] 

机构地区：[1]中山大学附属第三医院妇科,广东广州510630

出　　处：《中国病理生理杂志》2011年第9期1746-1750,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30772332);广东省科技计划资助项目(No.2009B060700028);广东省科技计划资助项目(No.2010B031600035);澳门科学技术发展基金资助项目(No.002/2009/A)

摘　　要：目的:研究CHFR在人子宫内膜癌细胞株中的表达和甲基化状态,以及去甲基化处理后对子宫内膜癌细胞克隆形成能力的影响。方法:半定量RT-PCR和Western blotting检测CHFR在子宫内膜癌细胞株的表达情况,甲基化特异性PCR检测CHFR的甲基化情况,用5-氨杂胞苷(5-aza)去甲基化处理RL-952细胞后再检测CHFR的表达和及其基因甲基化状态,平板克隆形成实验检测去甲基化对RL-952细胞的克隆形成能力的影响。结果:CHFR在子宫内膜癌细胞株中的表达均比正常成纤维细胞(NF)低,特别在RL-952细胞中表达显著降低。CHFR弱表达的RL-952细胞中CHFR异常高甲基化,而CHFR高表达的NF和ISH细胞均未见甲基化。用5-aza去甲基化处理RL-952细胞后,CHFR甲基化条带消失,其mRNA和蛋白表达明显增加,克隆形成率显著降低。结论:CHFR启动子甲基化是导致子宫内膜癌细胞中CHFR表达降低的主要原因,5-aza能恢复CHFR在子宫内膜癌细胞中的表达,并抑制癌细胞的克隆形成能力。AIM： To study the expression and methylation status of checkpoint with forkhead-associated and ring finger（CHFR） in human endometrial cancer cell lines and to explore the effect of demethylation on the colony formation of the cancer cells.METHODS： The expression level of CHFR in endometrial cancer cells was evaluated by semi-quantitative RT-PCR and Western blotting.The methylation status of CHFR was detected by methylation-specific PCR.The proliferation of RL-952 cells was analyzed by colony formation test.RESULTS： Lower expression of CHFR was detected in endometrial cancer cells,especially in RL-952 cells,than that in normal fibroblast（NF）.CHFR was weekly expressed in RL-952 cells,the gene of which had aberrant hypermethylation.The unmethylation band of CHFR was observed in NF cells and ISH cells,which had high expression of CHFR.After exposed to 5-azacytidine（5-aza）,the methylation band in RL-952 cells disappeared with the recovery of both CHFR mRNA and protein expression,and the colony-forming efficiency was greatly inhibited.CONCLUSION： Aberrant promoter methylation is the main reason for decreasing CHFR expression in endometrial cancer cells.5-aza could restore CHFR expression and inhibit the colony formation effectively.

关 键 词：子宫内膜肿廇 RL-952细胞 CHFR蛋白 人 甲基化 

分 类 号：R737.33[医药卫生—肿瘤]