乙型肝炎病毒X蛋白抑制SFRP5启动子的活性

发布时间：2018-01-03 17:27

本文关键词：乙型肝炎病毒X蛋白抑制SFRP5启动子的活性　出处：《中国病理生理杂志》2013年02期 　论文类型：期刊论文

【摘要】：目的:探讨乙型肝炎病毒X蛋白(HBx)抑制分泌型卷曲相关蛋白5(SFRP5)启动子活性的分子机制。方法:扩增不同长度的SFRP5启动子区片段,构建到萤光素酶报告基因载体pGL3-Basic上,并将构建好的重组质粒转染入正常肝细胞系LO2细胞,再分别感染编码HBx的腺病毒(Ad-HBx)或编码绿色荧光蛋白的腺病毒(Ad-GFP),荧光显微镜下观察病毒感染效率,检测萤光素酶活性以观察HBx对SFRP5启动子活性的影响。结果:(1)含不同长度的SFRP5启动子区截短突变报告质粒构建成功。(2)截短突变报告质粒的萤光素酶活性与pGL3-Basic对照组相比,分别为(6.32±0.04)倍(-478 bp～+47 bp)、(5.79±0.32)倍(-811 bp～+47 bp)、(3.59±0.34)倍(-1 235 bp～+47 bp)、(3.86±0.39)倍(-1 677 bp～+47 bp)、(3.26±0.42)倍(-2 072 bp～+47 bp)(均P0.05)。过表达HBx可以明显抑制SFRP5启动子活性,抑制率分别为44%(-478 bp～+47 bp)(P0.05)、46%(-811 bp～+47 bp)(P0.05)、28%(-1 235 bp～+47 bp)、24%(-1 677 bp～+47 bp)和40%(-2 072 bp～+47 bp)。结论:HBx可以明显抑制SFRP5启动子区活性,可能导致SFRP5基因表达下调。
[Abstract]:Objective: to investigate the inhibitory effect of hepatitis B virus X protein (HBX) on secretory crimp associated protein 5 (SFRP5). Methods: SFRP5 promoter fragments of different lengths were amplified. Luciferase reporter gene vector pGL3-Basic was constructed and the constructed recombinant plasmid was transfected into normal liver cell line LO2 cells. Then the adenovirus encoding HBx (Ad-HBx) or the adenovirus encoding green fluorescent protein (Ad-GFPX) were infected respectively. The infection efficiency was observed under fluorescence microscope. Luciferase activity was detected to observe the effect of HBx on the activity of SFRP5 promoter. The luciferase activity of truncated mutant reporter plasmid was compared with that of pGL3-Basic control group. They were 6.32 卤0.04) -478bp~ 47bp-579 卤0.32) -811bp-47bprespectively. (3. 59 卤0. 34) times (1 235 BP ~ 47 BP) and 3 86 卤0 39) times (1 677 BP ~ 47 BP). The overexpression of HBx could significantly inhibit the activity of SFRP5 promoter. The inhibition rates were -478bp~ 47bpP0.05 / 46 and -811bp-47bpP0. 05 respectively. 28 ~ 1 235 BP ~ 47 BP). 1 677 BP ~ 47 BP) and 40 ~ 2 072 BP ~ 47 BP. Conclusion the activity of SFRP5 promoter can be significantly inhibited by 1: HBx. This may lead to down-regulation of SFRP5 gene expression.
【作者单位】：重庆医科大学感染性疾病分子生物学教育部重点实验室;
【基金】：国家自然科学基金资助项目(No.31171307;o.30972586)
【分类号】：R373
【正文快照】： 全世界大约有4亿人感染乙型肝炎病毒(hepa-titis B virus,HBV),中国约占1/3[1]。乙型肝炎病毒属于嗜肝DNA病毒科,全长3.2 kb,含有4个部分双链的开放读码框,分别编码核心蛋白、表面抗原、X蛋白以及病毒DNA聚合酶,其中乙型肝炎病毒X蛋白(hepatitis B virus X protein,HBx)的生

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