摘要
目的研究Sirt5对人肝癌细胞系细胞增殖及迁移能力的影响及其机制。方法RT-PCR及Western Blot法检测Sirt5基因在正常肝细胞L02,HCC细胞系HepG2、Huh7、HepG3中的表达水平。HepG2及Huh7细胞过表达Sirt5,CCK-8法和细胞迁移实验检测Sirt5过表达对肝癌细胞增殖和迁移能力的影响。采用Western Blot法检测Sirt5过表达后HepG2细胞内STAT3蛋白活化水平和Nanog蛋白表达。用STAT3激活剂Colivelin预处理细胞1.5 h后过表达Sirt5,CCK-8法检测细胞增殖和迁移能力,并用Western Blot检测STAT3磷酸化水平。结果Sirt5在HepG2、Huh7、HepG33种细胞中均低表达,其中以HepG2细胞中表达水平最低。过表达Sirt5可抑制HepG2细胞增殖和迁移能力。Western Blot检测发现过表达Sirt5后可显著抑制STAT3的磷酸化和Nanog的水平,证明Sirt5可抑制STAT3/Nanog信号通路。且Sirt5抑制细胞增殖和迁移的能力可被STAT3激活剂Colivelin阻断。结论Sirt5可通过抑制STAT3/Nanog信号通路抑制HepG2细胞增殖。
Objective To study the effect and mechanism of Sirt5 on the proliferation and migration of human hepatocellular carcinoma cells.Methods The mRNA and protein expression of Sirt5 in different HCC cell lines HepG2,Huh7,HepG3 was detected.CCK 8 assay and cell migration assay were used to investigate the proliferation and migration ability of HepG2 cell.The expression of Nanog protein and activation of STAT3 protein in hepatocellular carcinoma HepG2 cell after overexpression Sirt5 were measured by Western Blot.The cells were pretreated with colivelin(STAT3 activator)for 1.5 h followed by treatment of Sirt5.CCK-8 assay was used to detect the proliferation and migration ability of HepG2.Moreover,the phosphorylation levels of STAT3 were detected by Western Blot.Results Sirt5 was downregulated in different HCC cell lines including HepG2,Huh7 and HepG3 when compared with L02.Overexpression of Sirt5 inhibited the cell viability and migration ability of HepG2 and Huh7 cells.Western Blot showed that Sirt5 overexpression significantly inhibited Nanog expression and phosphorylation of STAT3 protein.Moreover,the pro-proliferation effect of Sirt5 on HepG2 cell were blocked by colivelin,confirming that Sirt5 inhibits the STAT3/Nanog signaling pathway.Conclusion Sirt5 can inhibit the proliferation and migration ability of HepG2 cell by inhibiting STAT3/Nanog signaling pathway.
作者
肖金成
康鑫鑫
朱文良
白淇文
李靖
XIAO Jincheng;KANG Xinxin;ZHU Wenliang;BAI Qiwen;LI Jing(Department of Microinvasive Intervention,Tumour Hospital Affiliated to Zhengzhou University,Zhengzhou 450003,China;不详)
出处
《实用医学杂志》
CAS
北大核心
2019年第23期3593-3596,共4页
The Journal of Practical Medicine
基金
2018年河南省医学科技攻关计划项目（编号：2018020492）
