丙型肝炎病毒NS3/4A蛋白对人肝癌SMMC-7721细胞增殖、凋亡及分裂的影响  

Effects of hepatitis C virus NS3/4A protein on proliferation,apoptosis and division of human hepatocarcinoma SMMC-7721 cells

作　　者：李续亮[1] 景刚[2] Li Xuliang;Jing Gang(Department of Medical Laboratory Science,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China;Department of Medical Laboratory Science,the Second Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西省肿瘤医院中国医学科学院肿瘤医院山西医院山西医科大学附属肿瘤医院检验科,太原030013 [2]山西医科大学第二医院检验科,太原030001

出　　处：《肿瘤研究与临床》2022年第8期610-614,共5页Cancer Research and Clinic

摘　　要：目的探讨丙型肝炎病毒(HCV)NS3/4A蛋白对人肝癌SMMC-7721细胞增殖、凋亡及分裂的影响。方法采用同源重组方法构建重组质粒pLV-Green-NS3/4A和pLV-puro-NS3/4A, 将细胞分为pLV-Green-NS3/4A或pLV-puro-NS3/4A转染组及其相应对照组(转染空载体)。采用免疫荧光染色和蛋白质印迹法检测重组蛋白表达情况, 四甲基偶氮唑盐(MTT)法检测细胞增殖能力, DAPI染色法检测细胞凋亡情况, 免疫荧光分析细胞有丝分裂情况。结果 pLV-Green-NS3/4A转染组SMMC-7721细胞的生长状态差于相应空载体对照组;与相应空载体对照组比较, 第3、4天pLV-Green-NS3/4A转染组SMMC-7721细胞增殖活性均被抑制(均P<0.05)。pLV-Green-NS3/4A转染组和相应空载体对照组细胞凋亡率分别为(20.3±3.5)%和(5.3±1.5)%, 差异有统计学意义(t=-8.57, P<0.001)。与相应的空载体对照组比较, pLV-puro-NS3/4A转染组细胞出现更多的多极纺锤体, 两组多极分裂率分别为(2.33±0.58)%、(6.01±0.99)%, 差异有统计学意义(t=-5.50, P=0.005)。结论 HCV NS3/4A可抑制人肝癌SMMC-7721细胞增殖、促进细胞凋亡和多极分裂。Objective To investigate the effects of hepatitis C virus(HCV)NS3/4A protein on proliferation,apoptosis and division of human hepatocarcinoma SMMC-7721 cells.Methods The recombinant plasmid pLV-Green-NS3/4A and pLV-puro-NS3/4A were produced by homologous recombination method,and then the cells were divided into pLV-Green-NS3/4A or pLV-puro-NS3/4A transfection group and their corresponding control group(transfected with empty vector).The expression of recombinant protein was detected by immunofluorescence staining and Western blotting.Methyl thiazol tetrazolium(MTT)method was used to detect cell proliferation,DAPI staining was used to detect cell apoptosis,and immunofluorescence was used to analyze cell devision.Results The growth state of SMMC-7721 cells in the pLV-Green-NS3/4A transfection group was worse than that in the corresponding empty vector control group.Compared with the corresponding empty vector control group,the proliferation activity of SMMC-7721 cells in the pLV-Green-NS3/4A transfection group on the 3rd and 4th day was inhibited(both P<0.05).The apoptosis rates of cells in the pLV-Green-NS3/4A transfection group and the corresponding empty vector control group were(20.3±3.5)%and(5.3±1.5)%,and the difference was statistically significant(t=-8.57,P<0.001).Compared with the corresponding empty vector control group,the cells in the pLV-puro-NS3/4A transfection group had more multipolar spindles,and the multipolar division rates of the two groups were(2.33±0.58)%and(6.01±0.99)%,and the difference was statistically significant(t=-5.50,P=0.005).Conclusions HCV NS3/4A can inhibit the proliferation,promote the apoptosis and multipolar division of human hepatocarcinoma SMMC-7721 cells.

关 键 词：癌 肝细胞 肝炎病毒 丙型 细胞增殖 细胞凋亡 细胞分裂 

分 类 号：R735.7[医药卫生—肿瘤]