Function of interferon-γ induced HepG2 cell apoptosis via inhibiting autophagy
10.13699/j.cnki.1001-6821.2017.04.015
- VernacularTitle:干扰素γ抑制自噬诱导肝癌HepG2细胞凋亡的作用
- Author:
Shi-Peng LI
1
;
Zhen WANG
;
Jin-Dan HE
;
Yao YU
;
Hai-Ming ZHANG
;
Jian-Jun ZHANG
Author Information
1. 焦作市人民医院普外科
- Keywords:
interferon-γ;
hepatoma carcinoma cell;
autophagy;
apoptosis
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(4):343-346
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the mechanism of interferon-γ (IFN-γ) induced HepG2 cell apoptosis by inhibiting autophagy via Stat1-IRF1 pathway.Methods HepG2 cells were divided into control group (0) and experimental groups within 9 different doses of IFN-γ (10,20,30,50,75,100,150,200,300 ng · mL-1).HepG2 cells were treated with IFN-γfor 24 h.The cell viability of HepG2 cells was measured with MTT assay.Apoptotic cells were detected by Annexin V-FITC/propidium iodide (PI) kit.The quantity and distribution of LC3 dots in HepG2 cells were observed by fluorescence microscope.The expressions of phospho-signal transducer and activator of transcription-1 (p-Stat1),interferon regulatory factor-1 (IRF1),oxidative stress-activated poly (ADP-ribose) polymerase-1 (PARP1),autophagy regulation factor Beclin1 (Beclin1),autophagy gene Atg5 (Atg5),microtubule-associated protein 1 light chain 3 (LC3),cystein-asparate protease 3 (Caspase-3) and Caspase-8 were detected by Western blotting.Results As compared with control group (100％),IFN-γwith seven different doses (30,50,75,100,150,200,300 ng · mL-1),could suppress the proliferation of HepG2.The survival rates of these seven different doses of IFN-γ were (86.33 ± 4.72)％,(78.12±5.56)％,(66.33 ±5.13)％,(50.33 ±6.65)％,(46.07 ± 1.03)％,(37.67 ±7.05)％ and (31.33 ±3.51)％.The inhibitory effect significantly increased and the difference was statistically significant (P ＜0.01).As compared with that in the control group (3.77 ± 0.67)％,the apoptosis rate increased after the treatment with 100 ng · mL-1 IFN-γ which was (38.96 ±-4.35)％,and the difference between two groups was statistically significant (P ＜0.001).Moreover,there was a significant decrease of LC3 dots in 100 ng · mL-1 IFN-γ treated HepG2 cells 4.6 ± 1.1 as compared with that in the control group 15.6 ± 1.8,and the difference between two groups was statistically significant (P ＜ 0.001).HepG2 cells were treated with 100 ng · mL-1 IFN-γfor 24 h,then the expression of p-Stat1,IRF1,PARP1,Cleave Caspase-3 and Cleave Caspase-8 up-regulated,while the expression of Beclin1,Atg5,and LC3Ⅱdown-regulated.Conclusion IFN-γ could induce HepG2 cell apoptosis via inhibiting autophagy,which may be related to stimulating Stat1-IRF1 signaling pathway.
