HBxAg对胎盘滋养细胞凋亡的影响及其作用机制

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...

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Published in西安交通大学学报（医学版） Vol. 38; no. 6; pp. 809 - 814
Main Author 王慰敏 袁永兴 李琛 潘怡霞 徐娜 白桂芹
Format Journal Article
LanguageChinese
Published 西安交通大学第一附属医院妇产科,陕西西安,710061%西安交通大学第一附属医院基因联合实验室,陕西西安,710061 2017
Subjects
HTR-8
JEG-3
乙型肝炎病毒X
人胎盘滋养细胞
瞬时转染
细胞凋亡
细胞凋亡
JEG-3
乙型肝炎病毒X
瞬时转染
transient transfection
apoptosis
hepatitis B virus X
human placental tropho-blastic cell
人胎盘滋养细胞
HTR-8
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ISSN1671-8259
DOI10.7652/jdyxb201706006

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Summary: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。
Bibliography:WANG Wei-min1 , YUAN Yong-xing1 , LI Chen1 , PAN Yi-xia1, XU Na1 , BAI Gui-qin2 （1. Department of Obstetrics and Gynecology; 2. Gene Union Laboratory, the First Affiliated Hospital of Xi＇an Jiaotong University, Xi＇an 710061, China）
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 ear
ISSN:1671-8259
DOI:10.7652/jdyxb201706006