丙型肝炎病毒基因1b型F蛋白对人肝细胞癌细胞株凋亡的影响

杨静静; 张云; 邓小昭; 许可; 王忠灿; 王洁; 冯乐; 丁伟良

Abstract

杨静静,张云,邓小昭,许可,王忠灿,王洁,冯乐,丁伟良.丙型肝炎病毒基因1b型F蛋白对人肝细胞癌细胞株凋亡的影响[J].Chinese journal of Epidemiology,2009,30(4):388-392

丙型肝炎病毒基因1b型F蛋白对人肝细胞癌细胞株凋亡的影响

Influence of F protein of hepatitis C virus subtype 1b inhibits on human hepatocellular carcinom cell apoptosis

Received:September 22, 2008

DOI：

KeyWord: 丙型肝炎病毒 F蛋白凋亡肿瘤坏死因子α

English Key Word: Hepatitis C virus F protein Apoptosis Tumor necrosis factor α

FundProject:江苏省高校自然科学基金(03KJD330144)

Author Name	Affiliation
YANG Jing-fing	School of Public Health, Nanfing Medical University, Nanfing 210029, China
ZHANG Yun	School of Public Health, Nanfing Medical University, Nanfing 210029, China
DENG Xiao-zhao	南京军区军事医学研究所
XU Ke	江苏省疾病预防控制中心
WANG Zhong-can	南京军区军事医学研究所
WANG Jie	School of Public Health, Nanfing Medical University, Nanfing 210029, China
FENG Le	江苏省宜兴市人民医院
DING Wei-liang	江苏省宜兴市人民医院

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Abstract:

目的研究丙型肝炎病毒(HCV)基因1b型F蛋白对人肝细胞癌细胞株凋亡的影响。方法构建真核表达重组质粒pcDNA3.0-F-EGFP，并利用Lipofectamine2000转染人肝细胞癌细胞系，同时设pcDNA3.0-C-EGFP-阳性对照、pcDNA3.0-HcpG2为阴性对照及未转染为空白对照，以Act-D、TNFα诱导四组细胞系发生凋亡，AnnexinV-FITC/PI双染检测肿瘤细胞凋亡率，并使用细胞凋亡DNALadder检测，进一步验证HCV1b型F蛋白在肝细胞凋亡的生物学功能效应。结果pcDNA3.0-F-EGFP-细胞系在发生凋亡的时间上比peDNA3.0-C-EGFP-快，但相对空白质粒转染组及未转染细胞系有明显延迟，凋亡的发生率也明显低于阴性对照及空白对照细胞系(P<0.001)。结论外源性基因HCV1b型F的引入及表达，对细胞的凋亡有抑制作用。

English Abstract:

investigate the effects of F protein of hepatitis C virus subtype lb on the apoptosis of human hepatocellular carcinom cells.Methods cells were transfected with recombinant plasmid pcDNA3.0-F-EGFP and pcDNA3.0-F-EGFP- strain was exposed to Act-D and tumor necrosis factor a (TNFα) treatment in order to induce cell apoptosis with positive control pcDNA3.0-C-EGFP-,negative control pcDNA3.0-C-EGFP- and blank control.Annexin V-FITC/PI of flow cytometry was performed to determine the number of apoptotic cells.DNA Ladder was used to observe the isolation of apoptotic DNA fragments in the apoptotie cells.Results pcDNA3.0-F-EGFP- cell strain showed a much delayed apoptosis as well as obviously lowering the apoptotic rate when compared with the pcDNA3.0- strain and strain (P<0.001).Conclusion The introduction and expression of extraneous gene (the F gene of hepatitis C virus subtype 1b) could significantly inhibit the apoptosis of cells.

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