上调SMYD3对人胆管癌FRH0201细胞中DNMT3B表达及细胞增殖的影响

摘要

目的:研究人胆管癌细胞株FRH0201中SET和MYND结构域含有蛋白3(SET and MYND domain-containing protein 3,SMYD3)过度表达对DNA甲基化转移酶3B(DNA methyltransferase 3B,DNMT3B)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,RT-PCR检测细胞中DNMT3B mRNA水平的变化;Western blotting检测细胞中DNMT3B蛋白水平的变化;CCK-8检测细胞增殖能力的改变;流式细胞术检测细胞周期的改变.结果:以未处理组为对照,胆管癌细胞FRH0201在转染pEGFP-C3-SMYD3质粒后,DNMT3B蛋白及mRNA表达均显著上升(P＜0.01);细胞的增殖能力显著提高、细胞增殖速度加快(P＜0.05);进入G2/M期的细胞明显增多(P＜0.05).结论:过度表达SMYD3,可引起细胞中DNMT3B的表达上调并增强细胞增殖能力.%AIM:To explore ihe effect of SET and MYND domain - containing protein 3 ( SMYD3 ) over - expression on the expression of DNA methyltransferase 3B (DNMT3B) and the proliferation ability in human cholangiocarci-noma cell line FRH0201. METHODS; Transient transfection of SMYD3 eukaryotic expression plasmid pEGFP - C3 - SMYD3 into human cholangiocarcinoma cell line FRH0201 was performed. The expression of DNMT3B at mRNA and protein levels was detected by RT - PCR and Western blotting,respectively. Cell proliferation was examined by CCK -8 method and cell cycle situation was checked by flow cytometry. RESULTS; After transfected with SMYD3 eukaryotic expression plasmid pEGFP - C3 - SMYD3 , the over - expression of SMYD3 in FRH0201 cells was observed. Compared with the un-transfected cells, the expression of DNMT3B was significantly increased (P <0. 01) , the proliferation rate was obviously accelerated (P<0. 05) and the number of the cells in G2/M phase was significantly increased (P<0. 05) in FRH0201 cells transiently transfected with pEGFP - C3 - SMYD3 plasmid. CONCLUSION; The transient transfection of pEGFP -C3 - SMYD3 plasmid induces over - expression of DNMT3B and promotes the proliferation of human cholangiocarcinoma cell line FRH0201.