ADAR1通过RNA编辑上调ZNF655表达并促进人肝癌细胞系HepG2中HBV复制

基础医学与临床 ›› 2018, Vol. 38 ›› Issue (3): 312-316.

ADAR1通过RNA编辑上调ZNF655表达并促进人肝癌细胞系HepG2中HBV复制

李京源¹,李涛²,朱席琳³,伍晓盼³,刘英³

1. 北京协和医学院
2. 中国医学科学院基础医学研究所
3. 北京协和医学院基础学院；中国医学科学院基础医学研究所

收稿日期:2017-11-20 修回日期:2017-12-21 出版日期:2018-03-05 发布日期:2018-02-27
通讯作者: 刘英 E-mail:liuyingpumc@163.com
基金资助:
国家重点基础研究发展计划(973计划)

ADAR1 up-regulates ZNF655 expression via RNA editing and enhances HBV replication in HepG2 cell line

Received:2017-11-20 Revised:2017-12-21 Online:2018-03-05 Published:2018-02-27

摘要/Abstract

摘要： 目的探讨细胞中ADAR1对锌指蛋白ZNF655的作用及其对乙肝病毒（HBV）复制的影响。方法在人肝癌细胞系HepG2细胞中，利用桑格测序验证ZNF655 3’UTR存在ADAR1 RNA编辑位点；RT-qPCR检测ADAR1和ZNF655 mRNA的表达及HBV total RNA 和 HBV 3.5 kb RNA的表达；双荧光素酶报告基因检测荧光素酶的相对表达；Western blot 检测ADAR1和ZNF655蛋白的表达；ELISA检测ZNF655对HBV标志物HBsAg和HBeAg的影响。结果 ZNF655基因3’UTR上chr7:99575277位点在DNA水平为纯合型，在RNA水平为杂合型；ZNF655基因3’UTR上chr7:99575277位点的编辑型 G 比正常型 A 的荧光素酶活性显著升高( P < 0. 001)；在转录和翻译水平，ADAR1都显著增加了ZNF655的表达( P < 0. 001)；ZNF655 对HBV 的表达起到促进的作用。结论 ADAR1通过编辑ZNF655的3'UTR上的chr7:99575277位点，使编辑位点A转换成G，上调了基因的表达，进而起到对HBV复制的促进作用。

关键词: 作用于RNA的腺苷脱氨酶1, 锌指蛋白655, RNA 编辑, 乙肝病毒

Abstract: Objective To explore the effect of ADAR1 on ZNF655 and the regulation of ZNF655 on the expression of HBV. Methods Sanger sequencing was used to validate the 3 'UTR region of ZNF655 was edited by ADAR1. The expression of ADAR1 and ZNF655 mRNA as well as HBV RNA were detected by RT-qPCR. Dual luciferase report plasmid assay was used to detect the expression of luciferase. To detect the expression of ADAR1 and ZNF655 protein by Western blot. HBsAg and HBeAg was detected by ELISA. Results The chr7:99575277 loci on ZNF655 3 'UTR was homozygous in DNA level and hybrid in RNA level. On the 3'UTR editing site of ZNF655，the luciferase activity of the edited G allele was significantly higher than that of the normal A allele (P<0. 001). The expression of ZNF655 was upregulated by ADAR1 in the level of transcription and translation (P<0. 01).ZNF655 significantly promoted the expression of HBV. Conclusions The chr7:99575277 loci on ZNF655 3 'UTR is edited by ADAR1, promoting the the expression of ZNF655, which upregualated the the expression of HBV.

Key words: ADAR1, ZNF655, RNA editing, HBV

中图分类号:

R512.6+2

李京源李涛朱席琳伍晓盼刘英. ADAR1通过RNA编辑上调ZNF655表达并促进人肝癌细胞系HepG2中HBV复制[J]. 基础医学与临床, 2018, 38(3): 312-316.

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