摘要
目的探讨乙肝病毒X蛋白结合蛋白(HBXIP)对HepG2细胞周期和凋亡的影响。方法体外培养肝癌细胞HepG2,并以不转染细胞为对照组,以转染pCMV-HBX质粒的细胞为HBX组,以转染HBXIPsiRNA+pCMV-HBX质粒的细胞为HBXIP-i组(利用HBXIP siRNA干扰质粒沉默HepG2细胞中HBXIP基因表达)。Western blot检测HBX和HBXIP表达水平;克隆形成实验研究HBXIP对细胞增殖的影响;流式细胞仪检测HBXIP对细胞周期和细胞凋亡的影响。结果HBX组HBX蛋白表达上调,HBXIP-i组HBXIP蛋白表达下调。HBX组单细胞克隆数显著高于对照组和HBXIP-i组(P<0.01),HBXIP-i组单细胞克隆数显著高于对照组(P<0.01)。HBX组G0/G1期细胞显著高于对照组和HBXIP-i组(P<0.01),HBX组S期、G2/M期细胞显著低于对照组和HBXIP-i组(P<0.01)。HBX组细胞凋亡率显著低于对照组和HBXIP-i组(P<0.01),HBXIP-i组细胞凋亡率显著低于对照组(P<0.01)。结论HBXIP可促进肝癌细胞的增殖和抑制癌细胞凋亡。
Objective To investigate the effect of hepatitis B virus X protein interacting protein(HBXIP) on cell cycle and apoptosis of HepG2 cells and its possible mechanism.Methods HepG2 cells were cultured in vitro,and non-transfected cells as the control group,while cells transfected with pCMV-HBX plasmid as HBX group,and transfected with HBXIPsiRNA pCMV-HBX plasmid as HBXIP-i group(using HBXIP siRNA interference plasmid to silence HBXIP gene expression in HepG2 cells).The expression of HBX and HBXIP protein were detected by Western blot.The effect of HBXIP on cell proliferation was studied by clonogenesis,cell cycle and apoptosis by flow cytometry.Results HBX protein expression was up-regulated in HBX group,and HBXIP protein expression was down-regulated in HBXIP-i group.The number of single cell clones in HBX group was significantly higher than that in control group and HBXIP-i group(P<0.01),and that in HBXIP-i group was significantly higher than that in control group(P<0.01).Cells in G0/G1 phase of HBX group were significantly higher than those of control group and HBXIP-i group(P<0.01),while cells in S phase and G2/M phase of HBX group were significantly lower than those of control group and HBXIP-i group(P<0.01).The apoptosis rate of HBX group was significantly lower than that of control group and HBXIP-i group(P<0.01),and that of HBXIP-i group was significantly lower than that of control group(P<0.01).Conclusion HBXIP can promote the proliferation of hepatoma cells and inhibit the apoptosis of cancer cells.
作者
胡凯
刘成永
魏素梅
陆明
HU Kai;LIU Cheng-yong;WEI Su-mei;LU Ming(Department of Hepatology,Xuzhou Infectious Disease Hospital Affiliated to Xuzhou Medical University,Xuzhou,Jiangsu 221004,China;不详)
出处
《中国临床研究》
CAS
2020年第5期589-592,共4页
Chinese Journal of Clinical Research
基金
江苏省卫生计生委科研课题(H201506)。