基础医学与临床 ›› 2019, Vol. 39 ›› Issue (3): 360-364.

• 研究论文 • 上一篇    下一篇

蛋白激酶B通过调控低氧诱导因子-1表达干预人肝癌细胞系SMMC-7721增殖

许佐明1,谭川1,路远1,姚天尉1,吴贤建1,汪建初2,浦涧1   

  1. 1. 右江民族医学院
    2. 右江民族医学院附属医院肝胆外科/广西肝胆疾病临床医学研究中心
  • 收稿日期:2018-05-23 修回日期:2018-10-30 出版日期:2019-03-05 发布日期:2019-03-05
  • 通讯作者: 浦涧 E-mail:pujianym@163.com
  • 基金资助:
    广西医疗卫生适宜技术开发与推广应用项目

AKT intervention on the proliferation of human hepatoma cell line SMMC-7721 through regulating HIF-1 expression

  • Received:2018-05-23 Revised:2018-10-30 Online:2019-03-05 Published:2019-03-05

摘要: 目的 研究与分析敲低蛋白激酶B(PKB/AKT)对低氧诱导因子-1(HIF-1)表达及肝癌细胞增殖的影响。方法 随机将人肝癌细胞系SMMC-7721分设5组:空白对照(BC)组、阴性对照(siRNA-NC)组、HIF-1 siRNA转染(HIF1-siRNA)组、AKT siRNA转染(AKT-siRNA)组和HIF-1 AKT siRNA转染(HIF1-AKT-siRNA)组。用倒置荧光显微镜观察转染效率;CCK-8检测细胞增殖抑制率;Western blot检测HIF-1蛋白的表达。结果 转染效率均达到90%以上。与siRNA-NC组相比较,HIF1-siRNA组、AKT-siRNA组和HIF1-AKT-siRNA组的细胞增殖抑制率均升高,HIF-1蛋白表达水平均下降(P<0.05);与HIF1-AKT-siRNA组比较,HIF1-siRNA组和AKT-siRNA组的细胞增殖抑制率均下降,HIF-1蛋白表达水平均升高(P<0.05);与HIF1-siRNA组比较, AKT-siRNA组的细胞增殖抑制率下降,HIF-1蛋白表达水平升高(P<0.05)。各组HIF-1蛋白表达水平与细胞增殖抑制率存在负相关关系(r=-0.874, P<0.05)。结论 敲低HIF-1、AKT表达均可抑制人肝癌细胞系SMMC-7721增殖;HIF-1可能是肝癌细胞增殖的主要决定因子, HIF-1表达水平下调可抑制肝癌细胞增殖;敲低AKT表达可降低HIF-1表达水平,并协同HIF-1低表达的抑肝癌细胞增殖作用。

关键词: 关键词:SMMC-7721, 蛋白激酶B, 低氧诱导因子-1, siRNA, 细胞增殖

Abstract: Objective The effect of knockdown of AKT expression on the proliferation of hepatocellular carcinoma (HCC) and its regulation was analyzed. Methods SMMC-7721 Cell line was transfected with recombinant lentivirus contained a HIF-1 siRNA and/or AKT siRNA. Then, the transfection efficiency was observed by inverted fluorescence microscope, the cell growth inhibition rate was detected by cell counting kit-8 (CCK-8), the expression level of HIF-1 protein was detected by Western blot, and the protein expression of HIF-1 was analyzed statistically. Results SMMC-7721 cells was transfected with siRNA-lentivirus with ≥90% efficiency. The inhibition rate of HIF1-siRNA group, AKT-siRNA group, and HIF1-AKT-siRNA group decreased significantly as compared to the siRNA-NC group (P<0.05), and the expression of HIF-1 proteins was significantly higher. On the other hand, the inhibition rate of HIF1-AKT-siRNA group was significantly higher as compared to the HIF1-siRNA group and AKT-siRNA group, and the expression of HIF-1 proteins decreased significantly (P<0.05). while the inhibition rate in the HIF1-siRNA group was significantly higher as compared to the AKT-siRNA group, and the expression of HIF-1 proteins decreased significantly. There is a negative correlation between the expression of HIF-1 protein and the inhibition rate of cell proliferation (r=-0.874, P<0.05). Conclusion The downregulation of HIF-1 or AKT expression inhibits the cell proliferation in SMMC-7721 cells. HIF-1 plays a vital role in the cell proliferation in human hepatoma cell, down-regulation of HIF-1 protein expression can inhibit human hepatoma cell proliferation. The knockout of AKT resultes in downregulation of the protein expression of HIF-1 and intervenes on the proliferation of human hepatocellular carcinoma synergizing with down-regulation of HIF-1.

Key words: Key words: SMMC-7721, Akt, HIF-1, siRNA, cell proliferation