基础医学与临床 ›› 2016, Vol. 36 ›› Issue (7): 995-999.

• 研究论文 • 上一篇    下一篇

沉默Gli2基因对人肝癌细胞系SMMC-7721体外血管生成的影响

李建水1,吴斌2,严舒2,邓大炜1,曾丽娟1   

  1. 1. 川北医学院附属医院
    2. 川北医学院附属医院 肝胆外科
  • 收稿日期:2016-01-25 修回日期:2016-05-24 出版日期:2016-07-05 发布日期:2016-06-22
  • 通讯作者: 李建水 E-mail:q422528137@qq.com
  • 基金资助:
    四川省教育厅科研项目

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

  • Received:2016-01-25 Revised:2016-05-24 Online:2016-07-05 Published:2016-06-22

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qRT-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qRT-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。

关键词: Gli2, 肝癌, 血管生成, 侵袭

Abstract: Objective To explore the effect of Gli2 on the angiopoiesisin hepatpcellular carcinoma cell line SMMC-7721.Methods Complementary shRNA oligonucleotides targeting the Gli2 gene was designed and inserted into lentiviral vector.shRNA lentivirus were transfected intoSMMC-7721 cells and then fluorescence photographs were taken. The gene silencing efficiency was measured by qRT-PCR 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.Detection the expression of VEGF-A in supernant by ELISA;Meanwhile,the qRT-PCR、Western blot were used to examineVEGF-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 Gli2in the interfered groupwas lower,the capacity of angiopoiesis was significantly weaken,the expression of VEGF-A in supernant was decreased.meanwhile,the expression of VEGF-Aand MMP-2 was declined significantly(p<0.05). Conclusion Silencing Gli2 gene could suppress angiopoiesis ability in hepatpcellular carcinoma cell line SMMC-7721.its mechanism may be related to the down-regulation of VEGF-A and MMP-2.

Key words: Gli2, hepatocellular carcinoma, angiopoiesis, invasion

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