HBx蛋白羧基端30个氨基酸缺失对肝癌细胞生物学行为的影响  

Impact of mutant of hepatitis B virus X with C-terminal deletions of 30 amino acids on biological activity of hepatocellular carcinoma cells

作　　者：刘晓红[1] 朱明华[1] 曹晓哲[1] 郑建明[1] 陈颖[1] 

机构地区：[1]第二军医大学附属长海医院病理科,上海200433

出　　处：《肿瘤》2006年第3期232-235,共4页Tumor

基　　金：国家自然科学基金资助项目(编号:30070344;30070839);第二军医大学博士创新性课题项目资助(编号:013601401869)

摘　　要：目的:探讨羧基端缺失了30个氨基酸的乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)突变体ΔHBx和野生型HBx对肝癌细胞Huh7生物学行为的影响。方法:脂质体介导pcDNA3ΔHBx和pcDNA3HBx重组体转染人HBV(-)的肝癌细胞Huh7。PCR扩增Neo基因及Western blot检测转染重组体。运用细胞生长曲线绘制、平板克隆形成、流式细胞仪和裸鼠成瘤实验对转染pcDNA3ΔHBx、pcDNA3HBx和pcDNA3的细胞生物学活性进行检测。结果:pcDNA3ΔHBx组细胞生长速度较pcDNA3HBx组和pcDNA3组减慢,其倍增时间延长;pcDNA3ΔHBx组克隆形成率较pcDNA3HBx组和pcDNA3组下降;细胞周期检测显示pcDNA3ΔHBx表达使Huh7细胞G0/G1期→S期的进程明显减慢。结论:HBx羧基端缺失了30个氨基酸的突变体比野生型HBx具有明显抑制细胞增殖的能力,提示该区域对于HBx功能发挥起着至关重要的作用。Objective：To explore effects of mutant of hepatitis B virus （HBx） with C-terminal deletions of 30 amino acids on the biological activity of Huh7 hepatocellular carcinoma cells and compared with wildtype HBx. Methods：The mammal expression vector of pcDNA3ΔHBx and pcDNA3HBx were transfected into Huh7 human hepatoeellular carcinoma cells using liposome-mediation method. The transduction rate was determined by detecting the neo gene via PCR amplification. The expression of pcDNA3ΔHBx and pcDNA3HBx was detected by Western blot. Biological activity of pcDNA3ΔHBx and pcDNA3HBx-transfected Huh 7 cells was evaluated by examination of cell proliferation,clone formation,cell cycle,cell apoptosis, and tumorigenicity in nude mice. Results： MTT assay showed that the population growth rates of pcDNA3ΔHBx cells was slower than pcDNA3HBx and pcDNA3 cells （P〈0.05）. The doubling time of pcDNA3ΔHBx cell was significantly longer than that of pcDNA3HBx and pcDNA3 cells. Both clonogenicity and tumorigenicity in nude mice of pcDNA3ΔHBx cells were suppressed. FCM analysis showed that progression of pcDNA3ΔHBx cells from G0 to S phase was delayed. Conclusion： Compared with wildtype HBx, truncated HBx with deletion of 30 amino acid at C terminal has great ability to inhibit cell proliferation. It suggests that this segment is necessary for the function of HBx.

关 键 词：癌 肝细胞 肝炎病毒 乙型 突变/遗传学 HUH7细胞 

分 类 号：R735.7[医药卫生—肿瘤]