Tim-3在人肝癌细胞中的表达及其对细胞增殖与迁移能力的影响  

Expression of Tim-3 in hepatocellular carcinoma cells and its effect on cell proliferation and migration ability

作　　者：魏东[1] 匡怡 刘孟刚[1] 周波[1] 陈平[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所肝胆外科,重庆400042

出　　处：《重庆医学》2014年第16期1975-1978,1982,共5页Chongqing medicine

基　　金：国家自然科学基金面上资助项目(81270523)

摘　　要：目的研究T细胞免疫球蛋白及黏蛋白结构域分子-3(Tim-3)在人肝癌细胞系中的表达及其对肝癌细胞肿瘤生物学行为的影响。方法采用荧光实时定量PCR(FQ-PCR)和蛋白免疫印迹方法检测人正常肝细胞株L02和肝细胞肝癌(HCC)细胞株HepG2、SMMC7721中Tim-3在mRNA和蛋白质水平的表达。运用siRNA转染技术沉默Tim-3基因在HepG2细胞株中的表达,免疫印迹筛选干扰效率最高的基因片段。MTT和细胞划痕实验检测沉默Tim-3基因后对细胞增殖、迁移能力的影响。结果 FQ-PCR及免疫印迹方法检测显示,人HCC细胞株HepG2、SMMC7721中Tim-3mRNA和蛋白的表达均较正常肝细胞株L02明显增高(P<0.05)。转染Tim-3siRNA后,肝癌细胞株HepG2中Tim-3蛋白水平表达明显降低。与对照组比较,细胞的增殖、迁移能力显著下降。结论 Tim-3mRNA及蛋白水平在肝癌细胞株中表达明显升高,促进细胞的增殖、迁移。Objective To study the expression of T cell immunoglobulin mucin-3（Tim-3）in hepatocellular carcinoma（HCC）cell line and its influence on the oncobiological behavior of HCC cells.Methods The expression of Tim-3 mRNA and protein in human normal liver cell line L02 and HCC cell line HepG2 and SMMC7721 was assessed by FQ-PCR and Western blot.The siRNA Tim-3 fragments were designed to silence the Tim-3 gene expression in HepG2 cel1.HepG2 cells were transfected with siRNA by using LipofectamineTM 2000.The expression of Tim-3 protein was detected after transfection by Western blot to screen the effective siR-NA fragment.The proliferation and migration potential of hepG2 cells was evaluated after Tim-3 gene silence by the cell growth curve MTT assay and the wound healing assay.Results Both expressions of Tim-3 mRNA and protein in human HCC cell line HepG2 and SMMC7721 were significantly higher than those in normal liver cell line L02（P〈0.05）.After siRNA transfection,the protein expression of Tim-3 in experimental group was significantly lower than that of the control group.Compared with control group,the proliferative activity and the migration ability were decreased significantly.Conclusion Expressions of Tim-3 mRNA and protein are increased in HCC cell line.Tim-3 expression promotes HCC cell proliferation and migration.

关 键 词：癌 肝细胞 T细胞免疫球蛋白及黏蛋白结构域分子3 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]