Toll样受体4的表达及miR-TLR4干扰对HBV相关肝癌细胞生物学活性的影响  

Expression of Toll-like Receptor 4 and Impact of miR-TLR4 Interference on Biological Activity of HBV-related Hepatocellular Carcinoma Cells

作　　者：曾治民[1] 钟琼[1] 何静[1] 欧阳玉茗 肖震宇[1] 刘安文[2] 

机构地区：[1]赣州市人民医院肿瘤科,赣州341000 [2]南昌大学第二附属医院肿瘤科,南昌330006

出　　处：《肿瘤防治研究》2016年第2期116-122,共7页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金(81160305)

摘　　要：目的观察Toll样受体4在乙肝病毒相关肝癌细胞系中的表达及对其生物学活性的影响。方法Western blot检测Hep3B、Hep G2.2.15、Hep G2、SMMC7721及Huh7五种肝癌细胞中TLR4蛋白表达情况,选择TLR4表达高的HBV阳性肝癌细胞作为研究对象。构建4个mi R-TLR4质粒和一个阴性对照质粒,选择干扰效果明显的质粒转染TLR4表达高的HBV阳性肝癌细胞。实验分为正常组,阴性对照组,干扰质粒3组及干扰质粒4组。通过MTT法、克隆平板形成实验、流式细胞术分别检测干扰TLR4表达对肝癌细胞增殖、克隆形成、周期及凋亡的影响。结果 TLR4基因在五种肝癌细胞株中均有表达,在Hep3B细胞中表达最高。与正常组和阴性对照组相比,干扰TLR4表达后,Hep3B细胞的增殖能力明显受到抑制(P<0.05),克隆形成率显著下降(P<0.05),周期阻滞于G2/M期,并促进细胞凋亡(P<0.05)。结论 TLR4表达上调促进HBV相关肝癌细胞Hep3B生长增殖、周期再分布以及抑制凋亡,在肝癌发生发展过程中起重要作用。Objective To investigate the effects of toll-like receptor 4 expression on gene expression and cell growth of human HBV-related hepatocellular carcinoma（HCC） cells. Methods Toll-like receptor 4 protein expression were analyzed in five HCC cell lines, Hep3B, HepG2.2,15, HepG2, SMMC7721 and Huh7, by Western blot. The cells with TLR4 overexpression were selected to be further researched. Four groups, normal group, negative control group, plasmid No.3 group and plasmid No.4 group, were divided. After transfecting the HCC ceils with TLR4 overexpression via miR-TLR4 plasimids, cell proliferation was measured by MTT assay; cell clonogenieity was detected by clone formation assay; and cell apoptosis and cycle distribution were examined by flow cytometry. Results TLR4 was experssed in all cell lines, and it was the highest in Hep3B cells. Compared with the normal and negative control groups, interfering TLR4 expression could cause cell cycle block at G2/M phase, inhibit the proliferation and cloning efficiency of Hep3B, and promote apoptosis（P〈0.05）. Conclusion Up-regulation of TLR4 expression promotes the growth of HBV-related HCC cell Hep3B and cell cycle redistribution, and inhibits cell apotosis, which plays a key role in the carcinogenesis of HBV-related HCC.

关 键 词：TOLL样受体4 乙肝病毒 肝细胞癌 MI R-TLR4质粒 

分 类 号：R735.7[医药卫生—肿瘤]