逆转素对人肝癌细胞HepG2增殖、克隆形成及凋亡的影响  

Effect of reversine on cell proliferation, colony formation and apoptosis in human hepatocellular carcinoma cell HepG2

作　　者：朱玲玲 林涛发 谢丽平 王少扬 ZHU Ling-ling;LIN Tao-fa;XIE Li-ping;WANG Shao-yang(Dept of Infectious Diseases,The first Affiliated Hospital,Yijishan Hospital of Wannan Medical College,Wuhu Anhui 241001,China;Dept of Infectious Diseases,Dong＇fang Hospital of Xiamen University,Fuzhou 350001,China;Teaching-research Office of Infectious diseases,Fuzhou Genenral Hospital,Fuzhou 350001,China)

机构地区：[1]皖南医学院弋矶山医院感染科,安徽芜湖241001 [2]厦门大学附属东方医院感染科,福建福州350001 [3]福州总医院传染病教研室,福建福州350001

出　　处：《中国药理学通报》2018年第12期1740-1744,共5页Chinese Pharmacological Bulletin

基　　金：福州总医院创新团队项目(No 2014cxtd05)

摘　　要：目的探讨逆转素(reversine)对人肝癌细胞HepG2细胞增殖、克隆形成和凋亡的影响。方法人肝癌细胞HepG2经不同浓度reversine(0.1～1.6μmol·L^(-1))处理后,应用MTS法检测其对细胞增殖能力的影响;克隆形成实验评估reversine作用于肝癌细胞HepG2的远期效应;流式细胞术Annexin V-FITC/PI双染法检测reversine处理后肝癌细胞HepG2的凋亡率;Western blot法检测凋亡相关蛋白PARP、Bax、Bcl-2及Bcl-xL表达水平的变化。结果与对照组(DMSO)相比,不同浓度reversine对人肝癌细胞HepG2的增殖均有抑制作用(P<0.05),且呈剂量依赖关系,IC_(50)为0.94μmol·L-1。Reversine可以明显抑制Hep G2细胞的克隆形成能力(P <0. 05),且较高浓度reversine处理后,Hep G2细胞几乎不能形成肉眼可见的细胞克隆; reversine可以诱导Hep G2细胞凋亡(P <0. 05),且细胞的凋亡率与reversine的浓度呈正相关; Western blot结果显示,reversine孵育48 h后,Hep G2细胞中活化PARP、Bax的表达明显上调;同时,Bcl-2及Bcl-xL蛋白的表达下降。结论 Reversine可以明显抑制人肝癌细胞Hep G2细胞的增殖和克隆形成能力,并且可诱导Hep G2细胞的凋亡,其诱导凋亡与线粒体凋亡途径相关。Aim To investigate the suppressive effect of reversine on human hepatocellular carcinoma cell HepG2. Methods In this study, the antitumor effect of reversine was evaluated via cell proliferation assay, colony formation assay, apoptosis assay and Western blotting. Results Our data showed that reversine significantly suppressed the proliferation in HepG2 cell line in a time and dose-dependent manner （ P 〈0.05）. The IC 50 of reversine in HepG2 was 0.94μmol·L^ -1 . After incubation with reversine for 10 days, the colony formation of HepG2 was also inhibited. Furthermore, the macroscopic cell colony was virtually impossible to form with high-concentration of reversine treatment. In addition, our results showed that reversine could induce cell apoptosis in a dose-dependent manner （ P 〈0.05）. Moreover, using Western blot assay, the increased levels of cleaved-PARP and Bax were detected after incubation with reversine, and the expression level of Bcl-2 and Bcl-xL decreased in the meantime. Conclusion Reversine could effectively inhibit the cell proliferation and cell colony formation of human hepatocellular carcinoma cell HepG2 by inducing cell apoptosis via mitochondria-dependent pathway.

关 键 词：肝癌 增殖 克隆 凋亡 HEPG2 reversine 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]