HBxAg对胎盘滋养细胞凋亡的影响及其作用机制  

Effect of hepatitis B virus X on the apoptosis of placental trophoblastic cells and its potential mechanism

作　　者：王慰敏[1] 袁永兴[1] 李琛[1] 潘怡霞 徐娜 白桂芹[2] 

机构地区：[1]西安交通大学第一附属医院妇产科,陕西西安710061 [2]西安交通大学第一附属医院基因联合实验室,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2017年第6期809-814,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81370721);陕西省科技统筹创新计划(No.2016KTCL03-01);西安交通大学第一附属医院临床科研项目(No.XJTU1AHCR2014-018)~~

摘　　要：目的探讨乙型肝炎病毒X(HBx)蛋白对胎盘滋养层细胞凋亡水平的影响及可能的作用机制。方法将已构建好的HBx基因的真核表达载体pcDNA3.1(+)-HBx瞬时转染人绒毛膜癌细胞株JEG-3及HTR-8,分别以转染空载体pcDNA3.1(+)和未转染作为阴性对照组和空白对照组,转染后48h,以逆转录-聚合酶链式反应(RT-PCR)鉴定HBx-mRNA,间接免疫荧光法和蛋白印迹法(Western blot)鉴定HBxAg的表达;流式细胞仪AnnexinⅤ+PI双染方法检测细胞凋亡状态;间接免疫荧光法、流式细胞仪及Western blot检测PI3K、pAKT的表达。结果JEG-3及HTR-8转染组中均有HBx蛋白表达,而在对照组未被检测到;转染组JEG-3、HTR-8细胞早期凋亡率小于阴性对照组(P<0.05),PI3K和pAKT1的表达水平高于阴性对照组(P<0.05),然而AKT1蛋白表达水平在转染组、阴性对照组和空白对照组间无显著性差别。结论 HBx基因可以成功地转入JEG-3及HTR-8细胞,HBx转染后可抑制滋养细胞凋亡,这可能与HBx激活PI3K/pAKT信号通路有关。Objective To investigate the effect of hepatitis B virus X（HBx）protein on the apoptosis of placental trophoblastic cells and its potential mechanism.Methods A pcDNA3.1 expression vector of HBx gene was constructed and transfected into JEG-3 and HTR-8 human placental trophoblastic cell lines,respectively.After transfection for 48 h,RT-PCR and immunofluorescence analyses were made to detect HBx mRNA and protein expressions.Flow cytometry was used to detect the early apoptosis status of JEG-3 and HTR-8 cells.The expressions of PI3 K and p-Akt were detected by immunofluorescence and Western blotting.Results After transfection for48 h,RT-PCR and immunofluorescence analyses showed that HBx mRNA and protein expressions were detected in JEG-3 and HTR-8 cells.Flow cytometry revealed that early apoptosis of JEG-3 and HTR-8 cells was reduced by pcDNA-HBx transfection（P〈0.05）.Immunofluorescence and Western blotting showed that PI3 K and p-Akt were significantly upregulated in HTR-8 cells（P〈0.05）.Conclusion HBx gene can be transfected into JEG-3 and HTR-8 human placental trophoblastic cell lines,respectively.After the transfection,the early apoptosis of JEG-3and HTR-8 cells is reduced.Its inhibition on apoptosis is related to the activation of the PI3K/Akt signaling pathway.

关 键 词：JEG-3 HTR-8 乙型肝炎病毒X 瞬时转染 细胞凋亡 人胎盘滋养细胞 

分 类 号：R714.251[医药卫生—妇产科学]