沉默FATE/BJ-HCC-2基因的表达对肝癌细胞迁移及侵袭的影响  

Effects of silencing FATE/BJ-HCC-2 gene on migration and invasion of hepatocellular carcinoma cells

作　　者：葛丽丽[1,2] 朴军颜[1] 杨小昂[1] 尹艳慧[3] 张毓[3] 

机构地区：[1]郑州大学医药科学研究院,郑州450052 [2]郑州市儿童医院检验科,郑州450053 [3]北京大学免疫学系T细胞实验室,北京100083

出　　处：《郑州大学学报（医学版）》2015年第6期805-809,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目81071724;河南省基础与前沿基金资助项目142300413224

摘　　要：目的:研究FATE/BJ-HCC-2基因表达与肝癌细胞迁移及侵袭的关系。方法:体外合成FATE/BJ-HCC-2基因序列特异性小干扰RNA(siRNA),转染肝癌细胞系克隆株5B4,并设非特异性siRNA转染和空白对照组。采用RT-PCR及Western blot方法检测各组细胞中FATE/BJ-HCC-2 mRNA和蛋白的表达;通过Transwell小室模型检测各组细胞体外迁移及侵袭的能力;激光共聚焦显微镜下观察5B4及空质粒转染的对照细胞(Mock)的骨架结构。结果:各组细胞中FATE/BJ-HCC-2 mRNA和蛋白表达差异有统计学意义(F=15.321,P=0.020),特异性siRNA转染组细胞低于其他2组(P<0.05)。抑制FATE/BJ-HCC-2基因表达后,细胞体外迁移及侵袭的能力降低(F=6.171和10.109,P<0.05)。5B4细胞形态拉长,蝌蚪状,呈现迁移、游走的状态;微丝丰富,形态规则,平行贯穿于细胞全长,细胞膜周围有较多丝状伪足伸出;微管减少,分布不规则。结论:FATE/BJ-HCC-2基因表达促进肝癌细胞的迁移及侵袭。Aim：To study the effects of FATE/BJ-HCC-2 gene expression on the migration and invasion of hepatocel-lular carcinoma（HCC） cells.Methods：The HCC cell clone line 5B4 cells was transfected with FATE/BJ-HCC-2 gene se-quence-specific small interference RNA （ siRNA） synthesised in vitro .The expressions of FATE/BJ-HCC-2 mRNA and pro-tein were detected by RT-PCR and Western blot ,respectively .The migration and invasion ability of cells were examined by Transwell assay.The cytoskeleton of 5B4 cells transfected FATE/BJ-HCC-2 and empty vector control（Mock） cells was ob-served by laser scanning confocal microscope .The 5B4 cells not treated were used as blank control .Results：Both of mR-NA and protein expressions declined in the group of cells transfected with FATE /BJ-HCC-2 sequence-specific siRNA（F=15.321,P=0.020）.The migration and invasion ability of 5B4 cells obviously reduced after FATE/BJ-HCC-2 gene was si-lenced （F=6.171 and 10.109,P〈0.05）.Compared with that of the mock cells , the morphological and cytoskeleton structure of 5B4 cells transfected with FATE/BJ-HCC-2 changed obviously .Conclusion： FATE/BJ-HCC-2 gene expres-sion promotes migration and invasion of hepatocellular carcinoma cells .

关 键 词：FATE/BJ-HCC-2 RNA干扰 肝癌 转移 

分 类 号：R735.7[医药卫生—肿瘤]