流体动力学基因转染技术构建乙肝病毒HBx蛋白瞬时表达小鼠模型

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (5): 387-389,393.doi: 10.3969/j.issn.1004-616x.2017.05.013

• 技术与方法 • 上一篇下一篇

流体动力学基因转染技术构建乙肝病毒HBx蛋白瞬时表达小鼠模型

黄丹梅¹, 吴曼鹏², 黄晓华¹, 张莹¹, 倪仰鹏³, 牛永东¹

1. 汕头大学医学院药理教研室, 广东汕头 515041;
2. 汕头市第二人民医院儿科, 广东汕头 515011;
3. 揭阳市人民医院病理科, 广东揭阳 522091

收稿日期:2017-01-10 修回日期:2017-05-27 出版日期:2017-09-30 发布日期:2017-09-30
通讯作者: 牛永东,E-mail:ydniu@126.com E-mail:ydniu@126.com
作者简介:黄丹梅,E-mail:292710705@qq.com。
基金资助:
广东省科技计划项目（2014A 020212285）；汕头大学2015年“创新强校工程”专项资金建设项目（922-38040216）；广东省高水平大学重点学科建设项目（2015060，2016026，2015089）

Transient expression of HBx after hydrodynamic gene delivery in mice

HUANG Danmei¹, WU Manpeng², HUANG Xiaohua¹, ZHANG Ying¹, NI Yangpeng³, NIU Yongdong¹

1. Department of Pharmacology, Shantou University Medical College, Shantou 515041;
2. Department of Pediatrics, The Second People's Hospital of Shantou, Shantou 515011;
3. Department of Pathology, People's Hospital of Jieyang, Jieyang 522091, Guangdong, China

Received:2017-01-10 Revised:2017-05-27 Online:2017-09-30 Published:2017-09-30

摘要/Abstract

摘要： 目的：构建乙肝病毒HBx蛋白瞬时表达小鼠模型，为研究HBx在HBV感染相关原发性肝细胞癌发生中的分子机制奠定基础。方法：8只6~8周龄雄性CD-1小鼠，尾静脉注射真核表达质粒pcDNA-HBx及空载体pcDNA3.0，6~8 h处死小鼠并收集肝组织，抽提RNA及蛋白。RT-PCR和Western blot检测HBx表达情况；实时荧光定量PCR检测FXR靶基因SHP和Cyp7a1的mRNA表达。结果：核酸和蛋白水平上，pcDNA-HBx注射组小鼠肝脏内均可检测到HBx的表达；空载体对照组肝组织中未检测到HBx的表达；肝组织中瞬时过表达的HBx可以有效调控FXR通路下游靶基因SHP和Cyp7a1的mRNA表达。结论：成功实现了HBx在小鼠肝脏中的瞬时高表达，为今后整体水平上研究HBx提供了有效的动物模型。

关键词: 流体动力学, 基因瞬时表达, 尾静脉注射, HBx

Abstract: OBJECTIVE: To establish a transiently expressed HBx via hydrodynamic gene delivery in mice. METHODS: Two experimental groups of CD-1 male mice (4 mice per group) were injected with pcDNA-Flag-HBx plasmid or pcDNA empty vector via their tail veins. Mice were then sacrificed and their liver tissues were collected for extraction of RNA and protein. RESULTS: HBx was detected by PCR and Western blotting at the RNA and protein levels,respectively. Transiently expressed HBx effectively regulated the expression of typical target genes,small heterodimer partner (SHP) and cytochrome P450 7A1(Cyp7a1) of farnesoid X receptor signaling,as shown by real-time PCR analyses. CONCLUSION: A transient expression model of HBx via hydrodynamic gene delivery in mice was established.

Key words: hydrodynamic, transient gene expression, tail vein injection, HBx

中图分类号:

Q786

黄丹梅, 吴曼鹏, 黄晓华, 张莹, 倪仰鹏, 牛永东. 流体动力学基因转染技术构建乙肝病毒HBx蛋白瞬时表达小鼠模型[J]. 癌变·畸变·突变, 2017, 29(5): 387-389,393.

HUANG Danmei, WU Manpeng, HUANG Xiaohua, ZHANG Ying, NI Yangpeng, NIU Yongdong. Transient expression of HBx after hydrodynamic gene delivery in mice[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(5): 387-389,393.

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