沉默Gli2基因对人肝癌细胞系SMMC-7721体外血管生成的影响  

Effect of Gli2 gene silencing on the angiopoiesis of hepatocellular carcinoma cell line SMMC-7721

作　　者：李建水[1,2] 吴斌[2] 严舒[2] 邓大炜 曾丽娟[3] 

机构地区：[1]川北医学院肝胆胰肠研究所,四川南充637000 [2]川北医学院附属医院肝胆外科,四川南充637000 [3]川北医学院附属医院信息科,四川南充637000

出　　处：《基础医学与临床》2016年第7期995-999,共5页Basic and Clinical Medicine

基　　金：四川省教育厅科研项目(12ZB218)

摘　　要：目的探讨胶质瘤相关癌基因2(Gli2)对肝癌细胞SMMC-7721体外血管生成作用及机制。方法设干扰组、对照组和空白组;构建靶向Gli2的shRNA重组慢病毒和阴性对照慢病毒,转染到SMMC-7721细胞,RT-q PCR和Western blot检测沉默效率;用体外血管生成实验检测血管生成能力;用ELISA检测SMMC-7721细胞上清液中VEGF-A、MMP-2分泌水平;用RT-q PCR、Western blot检测细胞内VEGF-A、MMP-2的表达差异。结果 shRNA慢病毒转染率为90%左右,干扰组与对照组和空白组相比Gli2表达降低(P<0.05);血管生成能力减弱(P<0.05);上清液中VEGF-A含量下降(P<0.05);细胞中VEGF-A、MMP-2表达明显降低(P<0.05)。结论沉默Gli2的表达能够抑制肝癌细胞SMMC-7721血管生成,其机制可能与VEGF-A、MMP-2的下调有关。Objective To explore the effect of Gli2 on the angiopoiesis in hepatpcellular carcinoma cell line SMMC- 7721. Methods Complementary shRNA oligonucleotides targeting at the Gli2 gene was designed and inserted into lentiviral vector, shRNA lentivirus were transfeeted into SMMC-7721 cells and then fluorescence photographs were taken. The gene silencing efficiency was measured by RT-qPCR and Western blot. Interfered group, negative group and blank group were set up. Endothelial tube formation assay was used to detect the ability of angiopoiesis. Detec- tion the expression of VEGF-A in supernant by ELISA ; meanwhile, the RT-qPCR, Western blot were used to exam- ine VEGF-A and MMP-2 mRNA and protein expression. Results The transfection efficiency of shRNA lentivirus was about 90%. Compared with the negative group and blank group, the expression of Gli2 in the interfered group- was lower, the capacity of angiopoiesis was significantly weaken, the expression of VEGF-A in supemant was de- creased. Meanwhile,the expression of VEGF-A and MMP-2 declined significantly（P 〈0.05）. Conclusions Si- lencing Gli2 gene may suppress angiopoiesis in hepatpcellular carcinoma cell line SMMC-7721. Its mechanism may be related to the down-regulation of VEGF-A and MMP-2.

关 键 词：GLI2 肝癌 血管生成 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]