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...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24h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 eantigen，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HBVmRNA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。展开更多
AIM: p73, as a novel member of a family of p53-related transcription factors, shares redundant functions with p53, such as the abilities of inducing apoptosis and suppressing growth. It is well known that p53 can repr...AIM: p73, as a novel member of a family of p53-related transcription factors, shares redundant functions with p53, such as the abilities of inducing apoptosis and suppressing growth. It is well known that p53 can repress HBV expression and transcription efficiently. The aim of this paper is to investigate the transcriptional effect of p73α and p73β on hepatitis B virus (HBV) and to understand the correlation between HBV and p73.METHODS: To construct an x-gene inactivated HBV plasmid which was cotransfected with p73α or p73β expression vectors into HepG2 cells. After transiently transfecticn, HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) were detected by ELISA. Viral transcripts synthesized by HBV were evaluated by Northern blotting analysis. The activities of HBV regulatory elements, including enhancer Ⅰ/X promoter (ENI/Xp) and enhancer Ⅱ/core promoter (ENⅡ/Cp) were monitored by luciferase assays.RESULTS: Both p73α and p73β could repress HBsAg and HBeAg expression by downregulating the ENⅠ/Xp and ENⅡ/Cp activities. But p73β exerted stronger inhibition on the activity of ENI/Xp than p73α, resulting in much lower level of viral transcripts and the antigens expression.CONCLUSION: p73β as a novel member of p53 family can efficiently inhibit HBV transcription mainly through downregulating the activities of the HBV ENI/Xp regulatory elements.展开更多
乙肝病毒（Hepatitis B virus,HBV）是一种具有嗜肝性的DNA病毒,严重危害人类健康。据调查,15%～40%慢性乙型肝炎患者能逐渐进展成肝纤维化、肝衰竭,甚至肝癌。HBV基因组全长3.2kb,其中负链包含C、P、S和X等4个基因开放读码框（ORF）,...乙肝病毒（Hepatitis B virus,HBV）是一种具有嗜肝性的DNA病毒,严重危害人类健康。据调查,15%～40%慢性乙型肝炎患者能逐渐进展成肝纤维化、肝衰竭,甚至肝癌。HBV基因组全长3.2kb,其中负链包含C、P、S和X等4个基因开放读码框（ORF）,负责转录病毒各种mRNA。HBV mRNA转录在基因表达调控中起重大作用,受4个启动子（pre-C/C、Sp1、Sp2和X启动子）和两个增强子（enhancerⅠ和enhancerⅡ）正向调控。展开更多
文摘目的：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24h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 eantigen，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HBVmRNA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。
基金special funds for Major State Basic Research"973"of China,No.2001CB510205
文摘AIM: p73, as a novel member of a family of p53-related transcription factors, shares redundant functions with p53, such as the abilities of inducing apoptosis and suppressing growth. It is well known that p53 can repress HBV expression and transcription efficiently. The aim of this paper is to investigate the transcriptional effect of p73α and p73β on hepatitis B virus (HBV) and to understand the correlation between HBV and p73.METHODS: To construct an x-gene inactivated HBV plasmid which was cotransfected with p73α or p73β expression vectors into HepG2 cells. After transiently transfecticn, HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) were detected by ELISA. Viral transcripts synthesized by HBV were evaluated by Northern blotting analysis. The activities of HBV regulatory elements, including enhancer Ⅰ/X promoter (ENI/Xp) and enhancer Ⅱ/core promoter (ENⅡ/Cp) were monitored by luciferase assays.RESULTS: Both p73α and p73β could repress HBsAg and HBeAg expression by downregulating the ENⅠ/Xp and ENⅡ/Cp activities. But p73β exerted stronger inhibition on the activity of ENI/Xp than p73α, resulting in much lower level of viral transcripts and the antigens expression.CONCLUSION: p73β as a novel member of p53 family can efficiently inhibit HBV transcription mainly through downregulating the activities of the HBV ENI/Xp regulatory elements.
文摘乙肝病毒（Hepatitis B virus,HBV）是一种具有嗜肝性的DNA病毒,严重危害人类健康。据调查,15%～40%慢性乙型肝炎患者能逐渐进展成肝纤维化、肝衰竭,甚至肝癌。HBV基因组全长3.2kb,其中负链包含C、P、S和X等4个基因开放读码框（ORF）,负责转录病毒各种mRNA。HBV mRNA转录在基因表达调控中起重大作用,受4个启动子（pre-C/C、Sp1、Sp2和X启动子）和两个增强子（enhancerⅠ和enhancerⅡ）正向调控。
