基础医学与临床 ›› 2020, Vol. 40 ›› Issue (1): 37-40.

• 研究论文 • 上一篇    下一篇

巯基化Ras相关蛋白7a抑制人肝癌细胞系HepG2.2.15中HBV复制

宋佳茹, 朱席琳, 伍晓盼, 刘英*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 医学分子生物学国家重点实验室, 北京 100005
  • 收稿日期:2019-08-30 修回日期:2019-11-22 出版日期:2020-01-05 发布日期:2019-12-27
  • 通讯作者: *liuyingpumc@163.com
  • 基金资助:
    国家重点基础研究发展计划(973计划)(2012CB519005)

Sulfhydrated Ras related protein 7a inhibits HBV replication in human hepatocellular carcinoma cell line HepG2.2.15

SONG Jia-ru, ZHU Xi-lin, WU Xiao-pan, LIU Ying*   

  1. State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005, China
  • Received:2019-08-30 Revised:2019-11-22 Online:2020-01-05 Published:2019-12-27
  • Contact: *liuyingpumc@163.com

摘要: 目的 探究人肝癌细胞HepG2.2.15中Ras相关蛋白7a (Rab7a)的巯基化机制及其对乙型肝炎病毒(HBV)复制的影响。方法 通过生物素转换实验检测H2S对HepG2.2.15细胞中Rab7a巯基化水平的影响并验证Rab7a的巯基化位点;通过酶联免疫法、RT-qPCR和Western blot检测巯基化的Rab7a对HBV复制标志物水平的影响。结果 在HepG2.2.15细胞中H2S可增强内源性Rab7a的巯基化水平(P<0.01);NaHS(H2S速释供体)可增强外源性Rab7a的巯基化表达,但其巯基化位点突变后则不受NaHS影响(P<0.01);Rab7a巯基化后,可抑制HepG2.2.15细胞上清中HBsAg、HBeAg和HBV-DNA及细胞内HBV-cccDNA、3.5kb RNA、total RNA和HBcAg蛋白的表达(P<0.05)。结论 Rab7a的巯基化抑制HBV阳性细胞系HepG2.2.15细胞中HBV的复制水平。

关键词: 巯基化, 乙肝病毒, Ras相关蛋白7a

Abstract: Objective To investigate the mechanism of sulfhydrated Ras related protein 7a (Rab7a) in HepG2.2.15 cells and its effect on hepatitis B virus (HBV) replication. Methods The sulfhydrated Rab7a and sulfhydrated sites of Rab7a were verified by biotin switch method. The effect of the sulfhydrated Rab7a on the level of HBV replication markers was detected by enzyme-linked immunosorbent assay, RT-qPCR and Western blot. Results In HepG2.2.15 cells, H2S enhanced the sulfhydration level of endogenous Rab7a (P<0.01). NaHS (H2S rapid release donor) enhanced the expression of sulfhydration of exogenous Rab7a, but the mutation of sulfhydration site was not affected by NaHS (P<0.01). In HepG2.2.15 cells, H2S enhanced the sulfhydrylation level of endogenous and exogenous Rab7a; the sulfhydrylated Rab7a inhibited the expression of HBsAg, HBeAg and HBV-DNA in supernatant, as well as HBV-cccDNA, 3.5RNA, total RNA and HBcAg protein in HepG2.2.15 cells (P<0.05). Conclusions Sulfhydrated Rab7a inhibites the replication of HBV in HBV positive cell line HepG2.2.15.

Key words: sulfhydration, hepatitis B virus, Ras related protein 7a

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