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  

Transient expression of HBx after hydrodynamic gene delivery in mice

作　　者：黄丹梅[1] 吴曼鹏 黄晓华 张莹 倪仰鹏[3] 牛永东[1] HUANG Danmei;WU Manpeng;HUANG Xiaohua;ZHANG Ying;NI Yangpeng;NIU Yongdong(Department of Pharmacology, Shantou University Medical College,Shantou 515041;Department of Pediatrics, The Second People's Hospital of Shantou, Shantou 515011;Department of Pathology,People's Hospital of Jieyang, Jieyang 522091, Guangdong, China)

机构地区：[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

出　　处：《癌变．畸变．突变》2017年第5期387-389,393,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：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)

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pc DNA-HBx及空载体pc DNA3.0,6~8 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m RNA表达。结果:核酸和蛋白水平上,pc DNA-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m RNA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。OBJECTIVE:To establish a transiently expressed HBx via hydrodynamic gene delivery in mice.METHODS:Two experimental groups of CD-1male mice(4mice per group)were injected with pcDNA-Flag-HBx plasmidor p cDNA e mpty v ector v ia t heir t ail v eins.M ice w ere t hen s acrificed a nd t heir l iver t issues w ere c ollected f or e xtraction o f R NAand 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)andcytochrome P4507A1(Cyp7a1)of farnesoid X receptor signaling,as shown by real-time PCR analyses.CONCLUSION:Atransient e xpression m odel o f H Bx v ia h ydrodynamic g ene d elivery i n m ice w as e stablished.

关 键 词：流体动力学 基因瞬时表达 尾静脉注射 HBX 

分 类 号：Q786[生物学—分子生物学]