Screening host cell miRNA regulating HBV proliferation and HBsAg expression

章广玲; 王梅梅; 朱丽华; 熊亚南; 李怡璇; 刘民; 汤华; 袁丽杰

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Screening host cell miRNA regulating HBV proliferation and HBsAg expression

更新时间：2026-01-09

- Screening host cell miRNA regulating HBV proliferation and HBsAg expression
- Journal of Army Medical University Vol. 34, Issue 1, Pages: 44-48(2012)
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  CLC： R392-33;R373.21;R392.2
- Published：2012
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章广玲,王梅梅,朱丽华,熊亚南,李怡璇,刘民,汤华,袁丽杰.筛选调节HBV增殖和HBsAg产生的宿主细胞的miRNA[J].陆军军医大学学报(原第三军医大学学报),2012,34(01):44-48.

　Zhang Guangling,Wang Meimei,Zhu Lihua,Xiong Yanan,Li Yixuan,Liu Min,Tang Hua,Yuan Lijie.Screening host cell miRNA regulating HBV proliferation and HBsAg expression[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(01):44-48.
章广玲,王梅梅,朱丽华,熊亚南,李怡璇,刘民,汤华,袁丽杰.筛选调节HBV增殖和HBsAg产生的宿主细胞的miRNA[J].陆军军医大学学报(原第三军医大学学报),2012,34(01):44-48. DOI：

　Zhang Guangling,Wang Meimei,Zhu Lihua,Xiong Yanan,Li Yixuan,Liu Min,Tang Hua,Yuan Lijie.Screening host cell miRNA regulating HBV proliferation and HBsAg expression[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(01):44-48. DOI：

摘要

目的筛选调节乙型肝炎病毒（HBV）增殖和乙型肝炎病毒表面抗原（HBsAg）产生的宿主细胞的miRNA。方法用328种miRNA的反义链转染HepG2 2.2.15细胞，72 h收获上清，MTS检测细胞的增殖活性，ELISA检测表面抗原和e抗原的表达水平，实时定量PCR检测其中HBV DNA的拷贝数。构建筛选到的miRNA的过表达载体，转染HepG2 2.2.15细胞，也分别用MTS、ELISA和实时定量PCR进行检测。结果 miR-199a-3pASO、miR-210ASO和miR-185ASO促进了HBsAg产生和HBV增殖(P0.05);miR-199a-3p、miR-210、miR-185的过表达，分别抑制了HBsAg的产生和HBV的增殖(P0.05); miR-199a-3p、miR-210和miR-185的ASO转染组与对照组相比，分别降低了细胞内miR-199a-3p、miR-210和miR-185的表达水平(P0.05); miR-199a-3p、miR-210和miR-185 的过表达载体转染组与对照组相比，分别增加了细胞内miR-199a-3p、miR-210和miR-185的表达水平(P0.05)。结论在没有对细胞的增殖活性产生影响的情况下，miR-199a-3p、miR-210和miR-185抑制了HBV表面抗原的产生和HBV的增殖，miRNA作为病毒与宿主细胞相互作用中的调节因子，在HBV的生命活动周期中具有重要作用。

Abstract

Objective To screen out miRNAs that regulate HBsAg expression

HBeAg expression and proliferation of HBV in HepG2 2.2.15 cells

and to provide evidences for interaction between host cell’s miRNAs and HBV. Methods Three hundred and twenty eight antisense oligonucleotides of miRNAs were used to inhibit expressions of endogenous miRNAs. Enzyme linked immunosorbent assay (ELISA) and real-time PCR were performed on culture supernatant

which was harvested from HepG2 2.2.15 cells at 72 h after transfection with miRNA ASOs

to detect expressions of HBsAg and HBeAg and copy number of HBV DNA

respectively. Cell proliferation assay (MTS) was used at the same time. Results Three miRNAs

including miR-199a-3pASO

miR-210ASO and miR-185ASO

promoted HBsAg expression and HBV proliferation (P0.05). Overexpression of miR-199a-3p

miR-210 and miR-185 inhibited HBsAg expression and HBV proliferation (P0.05). Compared with those of the control group

HepG2 2.2.15 cells transfected with miR-199a-3pASO

miR-210ASO and miR-185ASO showed a decreased level of miR-199a-3p

miR-210 and miR-185

respectively (P0.05). Cells transfected with miR-199a-3p

miR-210 and miR-185 showed an increased levels of miR-199a-3p

miR-210 and miR-185 compared with those of the control group (P0.05). Conclusion miR-199a-3p

miR-210 and miR-185 inhibit HBsAg expression and HBV proliferation without inhibition of cell proliferation. miRNA

which regulates the interaction between host cell’s miRNAs and HBV replication

plays an important role in HBV proliferation.

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