过表达转录因子21抑制SMMC-7721肝癌细胞的增殖和迁移并促进其凋亡  

Over-expression of transcription factor 21 inhibits the proliferation and migration and promotes apoptosis of SMMC-7721 cells

作　　者：谭婧宇 张国庆[1] 刘锐[1] 周密[1] 李中堂[1] 吴忠均[1] 

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆400016

出　　处：《细胞与分子免疫学杂志》2015年第7期884-888,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：重庆市医学科研计划项目(2011-2-081)

摘　　要：目的探讨转录因子21(TCF21)基因对SMMC-7721肝癌细胞增殖、迁移和凋亡的影响及机制。方法采用脂质体将pc DNA3.1(+)TCF21及pc DNA3.1(+)质粒稳定转染入SMMC-7721细胞,分别为TCF21过表达组、空载体组、未处理组。采用反转录PCR检测转染前后TCF21 mRNA水平、Western blot法检测TCF21蛋白水平;MTT法检测细胞的增殖能力,划痕实验检测转染后细胞的迁移能力;异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测细胞凋亡率,Western blot法检测转染后KISS1、P53和基质金属蛋白酶9(MMP-9)的蛋白水平。结果 TCF21基因转染SMMC-7721细胞后,细胞内TCF21 mRNA和蛋白水平均明显增加。与对照细胞相比,过表达TCF21的肝癌细胞增殖能力减弱、癌细胞迁移能力降低、细胞凋亡增加。Western blot结果显示上调TCF21表达,可增加KISS1、P53水平并降低MMP-9的水平。结论过表达TCF21可抑制SMMC-7721肝癌细胞的增殖、迁移并促进其凋亡。Objective To explore the impact of transcription factor 21 （TCF21） gene on proliferation, migration and apoptosis in SMMC-7721 hepatocellular carcinoma cell line and the related mechanism. Methods Using Lipofectamine^TM 2000, we stably transfected pcDNA3.1 （＋） TCF21 and pcDNA3.1 （＋） plasmids into SMMC-7721 cells of TCF21 over-expression group and empty vector group, respectively. The untreated cells were set as blank control group. The expression of TCF21 mRNA and protein were investigated by reverse transcription PCR and Western blotting. Cell proliferation ability was detected by MIF assay. Wound healing assay was used to observe cell migration ability. Annexin V-FITC/PI double labeling combined with flow cytometry was applied to determine cell apoptosis rate. Western blotting was performed to examine the expressions of KISS1, P53 and matrix metalloproteinase-9 （MMP-9）. Results TCF21 was over-expressed in TCF21-transfected SMMC-7721 cells. Compared with the control groups, the TCF21 over-expression group showed relatively weakened proliferation ability and significantly inhibited migration ability as well as the increased apoptosis. Western blotting demonstrated that up-regulated TCF21 raised the expressions of KISS1 and 1353, and down-regulated MMP-9 level. Conclusion Tumor-suppressor gene TCF21 could inhibit the proliferation and migration of SMMC-7721 hepatocellular carcinoma cells and promote its apoptosis.

关 键 词：转录因子21(TCF21) 肝细胞肝癌 增殖 迁移 凋亡 

分 类 号：R735.7[医药卫生—肿瘤] Q279[医药卫生—临床医学]