丙型肝炎病毒核心蛋白可抑制人肝细胞的凋亡并促进其自噬。

HCV core protein represses the apoptosis and improves the autophagy of human hepatocytes.

作者信息

Liu Changhong, Qu Aihua, Han Xiaochun, Wang Yiguo

机构信息

School of Medicine, Shandong University Jinan 250012, China ; Department of Gastroenterology, Shandong Provincial Qianfoshan Hospital Jinan 250014, China.

Zouping Affiliated Hospital of Taishan Medical College Zouping 256200, China.

出版信息

Int J Clin Exp Med. 2015 Sep 15;8(9):15787-93. eCollection 2015.

PMID:26629077

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4658966/

Abstract

OBJECTIVES

This study aims to investigate the influence on human hepatocytes apoptosis and autophagy by the hepatitis C virus (HCV) core protein.

METHODS

QSG-7701, a human-derived non-neoplastic liver cell line, was transfected with PIRES-core vector that was a eukaryotic vector to express HCV core protein. Fluorescence microscope was used to observe the changes of nuclei in apoptosis cells by Annex in V-FITC/PI double staining. Flow cytometry was applied to detect the rate of cell apoptosis. Western blotting was used to detect the expression of HCV core protein, transcription factor nuclear factor-kappa B (NF-κB), autophagic biomarker microtubule associated protein 1 light chain 3 (LC3), and Beclin-1.

RESULTS

The apoptosis rate was significantly lower (P < 0.05) in QSG7701/core group (transfected with PIRES-core vector, (1.34±0.07)%) than in QSG7701 group (no transfection, (2.35±0.11)%) and in QSG7701 QSG7701/pcDNA3.1 group (transfected with pcDNA3.1 vector, (2.58±0.1)%). NF-κB expression was up-expressed in QSG7701/core group than in QSG7701/pcDNA3.1 group and QSG7701 group (P < 0.05). LC3-II expression and Beclin-1 expression was significant higher in QSG7701/core group than in the QSG7701/pcDNA3.1 group and QSG7701 group (P < 0.05).

CONCLUSION

HCV core protein can repress the apoptosis and improve the autophagy of QSG7701 through up-regulating NF-κB and Beclin-1 expression.

摘要

目的

本研究旨在探讨丙型肝炎病毒（HCV）核心蛋白对人肝细胞凋亡和自噬的影响。

方法

将真核表达载体PIRES-core转染人源非肿瘤性肝细胞系QSG-7701，以表达HCV核心蛋白。采用Annexin V-FITC/PI双染法，通过荧光显微镜观察凋亡细胞的核变化。应用流式细胞术检测细胞凋亡率。采用蛋白质免疫印迹法检测HCV核心蛋白、转录因子核因子κB（NF-κB）、自噬生物标志物微管相关蛋白1轻链3（LC3）和Beclin-1的表达。

结果

QSG7701/core组（转染PIRES-core载体，凋亡率为（1.34±0.07）%）的凋亡率显著低于QSG7701组（未转染，凋亡率为（2.35±0.11）%）和QSG7701/QSG7701/pcDNA3.1组（转染pcDNA3.1载体，凋亡率为（2.58±0.1）%）（P<0.05）。QSG7701/core组的NF-κB表达高于QSG7701/pcDNA3.1组和QSG7701组（P<0.05）。QSG7701/core组的LC3-II表达和Beclin-1表达显著高于QSG7701/pcDNA3.1组和QSG7701组（P<0.05）。