Liu Changhong, Qu Aihua, Han Xiaochun, Wang Yiguo
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.
Int J Clin Exp Med. 2015 Sep 15;8(9):15787-93. eCollection 2015.
This study aims to investigate the influence on human hepatocytes apoptosis and autophagy by the hepatitis C virus (HCV) core protein.
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.
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).
HCV core protein can repress the apoptosis and improve the autophagy of QSG7701 through up-regulating NF-κB and Beclin-1 expression.
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。
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。
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）。
HCV核心蛋白可通过上调NF-κB和Beclin-1表达抑制QSG7701细胞凋亡并促进自噬。
