引用本文:祝艺耘,付 愚,李中堂,刘 锐,魏续福,吴忠均.设计特异性结合HBV X基因启动子的人工锌指蛋白以抑制HepG2.2.15细胞中HBV的转录[J].重庆医科大学学报,2014,38(7):929~932
设计特异性结合HBV X基因启动子的人工锌指蛋白以抑制HepG2.2.15细胞中HBV的转录
Inhibition of HBV transcription in HepG2.2.15 cells by artificial Zinc finger protein designed specifically to target HBX gene promoter
DOI:
中文关键词:  乙型肝炎病毒  X基因启动子  锌指蛋白  HepG2.2.15细胞
英文关键词:hepatitis B virus  X gene promoter  Zinc finger protein  HepG2.2.15 cell
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祝艺耘,付 愚,李中堂,刘 锐,魏续福,吴忠均 重庆医科大学附属第一医院肝胆外科重庆 400016 
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中文摘要:
      目的:设计人工锌指蛋白(zinc finger protein,ZFP)特异性结合于乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBX)基因启动子,观察ZFP在体外对乙型肝炎病毒(hepatitis B virus,HBV)转录的抑制。方法:将pcDNA3.1-ZFP转染入HepG2.2.15细胞24 h后,用Western blot检测HBX蛋白的含量;用ELISA和real-time PCR检测上清液中乙型肝炎核心相关抗原(hepatitis B e antigen,HBeAg)和HBV DNA含量,用RT-PCR检测胞内HBV mRNA水平。结果:ZFP可抑制HepG2.2.15细胞中HBX的表达;相比空质粒组,ZFP组细胞培养上清液中HBV DNA 拷贝量和HBeAg 含量分别下降了51.7%(t=23.079,P=0.000,95%CI=44.98%~58.52%)、33.8%(t=3.887,P=0.003,95%CI=12.12%~55.48%);细胞内HBV mRNA也明显减少(t=3.616,P=0.022)。结论:人工设计的可特异性结合于HBX的ZFP可于HepG2.2.15细胞中有效抑制HBV转录。
英文摘要:
      Objective:To observe the inhibition of hepatitis B virus(HBV) transcription in vitro by Zinc finger protein(ZFP) which was designed specifically to target X gene promoter of HBV(HBX). Methods:Recombinant plasmid pcDNA3.1-ZFP was transfected into HepG2.2.15 cells. Expression of HBX protein was detected by Western blot at 24 h after being transfected. Hepatitis B e antigen(HBeAg) and HBV DNA levels in supernatant of HepG2.2.15cells at 24 h after being transfected with pcDNA3.1-ZFP were detected by ELISA and real-time PCR. HBV mRNA was tested by RT-PCR. Results:ZFP could inhibit the expression of HBX in HepG2.2.15 cells. In the presence of ZFP,HBV DNA level and HBeAg was considerably reduced by 51.7%(t=23.079,P=0.000,95%CI=44.98%-58.52%) and 33.8%(t=3.887,P=0.003,95%CI=12.12%-55.48%) respectively,while HBV mRNA was sharply decreased(t=3.616,P=0.022). Conclusion:ZFP protein designed to target X gene promoter of HBV can inhibit the transcription of HBV in HepG2.2.15 cells effectively.
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