HBVDNA聚合酶反式激活蛋白1反式下调CDC42结合蛋白4基因的表达

CDC42-interacting protein 4 gene is down trans-regulated by HBV DNA polymerase trans-activated protein 1

目的对乙型肝炎病毒(HBV)DNA聚合酶反式激活蛋白1(HBVDNAPTP1)的生物活性进行检测。方法构建pcDNA3.1(-)/myc-His A-HBVDNAPTP1载体,并使用该载体对单核细胞性白血病细胞系THP-1进行转染。通过Western blot蛋白印迹法对HBVDNAPTP1表达进行检测。使用抑制消减杂交技术(SSH)在pGEM-T Easy系统中构建THP-1细胞HBVDNAPTP1反式激活cDNA基因文库。在对cDNA序列进行测序后,将测序结果与GenBank中的序列进行BLAST对比检测分析。结果某些序列(如CIP4)可能参与细胞凋亡过程。THP-1细胞内的CIP4mRNA和蛋白表达分别通过实时定量反转录聚合酶链式反应(RT-PCR)和Western blot蛋白印迹法进行分析。HBVDNAPTP1能够在转录水平和翻译水平使CIP4的表达下调。结论 HBVDNAPTP1可能参与单核细胞凋亡起始的正向调节。本实验研究结果揭示了HBVDNAPTP1的生物功能,并为HBVDNAPTP1的调节机制的进一步探索提供了新证据。

Objective To determine the biological function of Hepatitis B Virus（HBV）DNA polymerase transactivated protein 1（HBVDNAPTP1）.Methods The vector pcDNA3.1（-）/myc-His A-HBVDNAPTP1 was constructed and used to transfect acute monocytic leukemia cell line THP-1.The expression of HBVDNAPTP1 was detected with western blot analysis.A cDNA library of genes transactivated by HBVDNAPTP1 in THP-1cells was made in pGEM-T Easy using suppression subtractive hybridization（SSH）.The cDNAs were sequenced and analyzed with BLAST search against the sequences in GenBank.Results Some sequences,such as CIP4,were found to be possibly involved in apoptosis.The expressions of CIP4 mRNA and protein were identified by Real time RT-PCR and western blot in THP-1cells.HBVDNAPTP1 was able to down-regulate the expression of CIP4 at both transcription and translation levels.Conclusion HBVDNAPTP1 may be involved in the positive regulation on the initiation of monocyte apoptosis.The result contributed to reveal the biological functions of HBVDNAPTP1 and provided new evidences for further exploration of the regulatory mechanism of HBVDNAPTP1.