SMYD3过表达对人肝内胆管癌细胞miR-124表达及细胞增殖的影响  

Effect of SMYD3 over-expression on miR-124 expression and proliferation ability in human intrahepatic cholangiocarcinoma cells

作　　者：郑礼平[1] 李志花[1] 陈汝福[2] 曾兵[2] 周嘉嘉[2] 龚远锋[2] 宋亚东[1] 

机构地区：[1]中山大学孙逸仙纪念医院 肿瘤科 [2]中山大学孙逸仙纪念医院 肝胆外科,广东广州510120

出　　处：《中国病理生理杂志》2013年第1期76-80,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30872485)

摘　　要：目的:探讨肝内胆管癌细胞HCCC-9810中SET和MYND结构域含有蛋白3(SET and MYND domain-containing protein 3,SMYD3)过表达对miR-124表达及细胞增殖能力的影响。方法:瞬时转染SMYD3真核表达质粒后,Western blotting检测细胞中SMYD3蛋白水平的变化;qRT-PCR检测细胞中SMYD3 mRNA和miR-124表达;甲基化特异性PCR检测miR-124-1、miR-124-2和miR-124-3基因启动子甲基化水平;CCK-8和平板克隆形成实验检测细胞增殖能力。结果:以空白组为对照,肝内胆管癌细胞HCCC-9810在转染pEGFP-SMYD3质粒后,SMYD3蛋白及mRNA表达均显著上升(P<0.05),miR-124-1、miR-124-2和miR-124-3基因启动子甲基化显著增加(P<0.05),miR-124表达明显下降(P<0.05),细胞增殖能力显著提高(P<0.05)。结论:过表达SMYD3可引起miR-124基因启动子甲基化增加,导致胆管癌细胞中miR-124的表达下降,同时过表达SMYD3可增强细胞增殖能力。AIM ： To explore the effect of SET and MYND domain-containing protein 3 （SMYD3） over-expres- sion on miR-124 expression and proliferation ability of human intrahepatic cholangiocarcinoma cells. METHODS： Transi- ent transfection of SMYD3 eukaryotic expression plasmid into human intrahepatic cholangiocarcinoma cell line HCCC-9810 were performed. The expression of SMYD3 at mRNA and protein levels was measured by qRT-PCR and Western blotting, respectively. The expression of miR-124 was detected by qRT-PCR, and the methylation status of miR-124 gene was deter- mined by methylation-specific PCR. Cell proliferation was examined by CCK-8 assay and colony formation experiment. RE- SULTS： After transfected with SMYD3 eukaryotic expression plasmid, the over-expression of SMYD3 in HCCC-9810 cells was observed. Compared with the blank ceils, the expression level of miR-124 was significantly decreased and miR-124 gene promoter methylation was significantly increased. In addition, SMYD3 over-expression significantly promoted the pro- liferation of HCCC-9810 cells. CONCLUSION： The transient transfection of SMYD3 plasmid increases the methylation of miR-124 gene promoter and induces under-expression of miR-124. Over-expression of SMYD3 promotes the proliferation of cholangiocarcinoma cells.

关 键 词：HCCC-9810细胞 SET和MYND结构域含有蛋白3 miR-124 甲基化特异性PCR 细胞增殖 

分 类 号：R735.8[医药卫生—肿瘤]