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

摘要

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.%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-3 mRNA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-3 siRNA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-3 mRNA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。