乙肝病毒X 蛋白对正常肝细胞生物钟基因表达的影响
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杨盛力, Email: jixufendou2013@gmail.com

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国家自然科学基金资助项目(81272421)。


Effects of hepatitis B virus X protein on clock gene expression in normal hepatic cells
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    摘要:

    目的:探讨乙肝病毒X蛋白(HBx)对人正常肝细胞中生物钟基因表达的影响。 方法:分别将HBx表达质粒(pcDNA3.1-HBx)与空载体质粒(pcDNA3.1)转染入人正常肝细胞系L02后,用RT-PCR与Western blot法检测细胞HBx mRNA与蛋白的表达;real-time PCR检测细胞中生物钟基因CLOCK、BMAL1、Per1、Per2、Per3、Cry1、Cry2、CKIε的表达。 结果:RT-PCR与Western blot结果显示,L02细胞转染HBx表达质粒后有明显的HBx mRNA与蛋白表达,而转染空载体质粒的L02细胞无HBx mRNA与蛋白表达。与转染空载体质粒的L02细胞比较,转染HBx表达质粒的L02细胞CLOCK与Cry1的mRNA表达明显升高,而BMAL1、Per1、Per2、Per3、Cry2、CKIε的mRNA表达明显降低,差异均有统计学意义(均P<0.05)。 结论:HBx能使正常肝细胞中的生物钟基因表达发生改变,破坏肝细胞正常的生物节律,这可能为其导致肝癌形成的机制之一。

    Abstract:

    Objective: To investigate the influence of hepatitis B virus X protein (HBx) on the clock gene expression in normal hepatic cells. Methods: Human normal hepatic L02 cells were transfected with the HBx expression plasmid (pcDNA3.1-HBx) or empty vector plasmid (pcDNA3.1), respectively, then the HBx mRNA and protein expression in L02 cells was determined by RT-PCR method and Western blot analysis, respectively, and the mRNA expression of the clock genes that included CLOCK, BMAL1, Per1, Per2, Per3, Cry1, Cry2 and CKIε in L02 cells were detected by real-time PCR. Results: Results of RT-PCR and Western blot showed that both HBx mRNA and protein were obviously expressed in the L02 cells transfected with HBx expression plasmid, which were not seen in the L02 cells transfected with empty vector plasmid. Compared with L02 cells transfected with empty vector plasmid, the CLOCK and Cry1 mRNA expressions were remarkably increased while the mRNA expressions of BMAL1, Per1, Per2, Per3, Cry2 and CKIε were remarkably decreased in L02 cells transfected with empty vector plasmid, and all the differences had statistical significance (all P<0.05). Conclusion: HBx can change the expressions of clock genes in normal hepatic cells, which may disrupt circadian rhythm of liver cells, and this may be one of the mechanisms for the pathogenesis of liver cancer.

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金涛,刘平,黄早早,李常海,姚弘毅,柳湘洁,杨盛力.乙肝病毒X 蛋白对正常肝细胞生物钟基因表达的影响[J].中国普通外科杂志,2014,23(7):915-920.
DOI:10.7659/j. issn.1005-6947.2014.07.010

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  • 收稿日期:2014-03-22
  • 最后修改日期:2014-06-08
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  • 在线发布日期: 2014-07-20