Effects of Gecko peptide mixture on HepG2 cells proliferation and endoplasmic reticulum stress pathway
10.13699/j.cnki.1001-6821.2018.02.016
- VernacularTitle:壁虎多肽混合物对HepG2细胞增殖及内质网应激途径的影响
- Author:
Yi-Meng DUAN
1
;
Leng-Xin DUAN
;
Ling LIU
;
Meng-Li GUO
;
Bing-Bing WANG
;
Ze-Yue HUANG
;
Jian-Gang WANG
Author Information
1. 河南科技大学医学院药理学与分子生物学实验室
- Keywords:
Gecko polypeptide mixture;
HepG2 cell;
endoplasmic reticulum stress;
C/EBP-homologous protein
- From:
The Chinese Journal of Clinical Pharmacology
2018;34(2):148-151
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Gecko polypeptide mixture (GPM) on the proliferation and endoplasmic reticulum stress (ERS) pathway of human hepatocellular carcinoma HepG2 cells.Methods The HepG2 cells were treated with differentconcentration of GPM(0,0.15,0.20,0.25,0.30,0.35,0.40,0.45 mg · mL-1) for 24 h,and then corresponding indicators were detected with respective methods.The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTF) assay was used to detect the viability of HepG2 cells.The concentration of blank group,GPM low-dose,middle-dose and high-dose experimental groups was respectively 0,0.1,0.2,0.3 mg · mL-1,according to the results of MTT.5-Fluorouracil was chosen as the positive control drug,which concentration was 10 μg · mL-1.Western blot analysis was applied to observe the expression of ERS-related proteins and apoptosis-related proteins in HepG2 cells.Results The GPM could inhibit the proliferation of HepG2 cells in a dose-and time-dependent manners.After treatment of GPM for 24,48,72 h,the 50％ inhibitory dose (IC50) values were 0.27,0.23,0.20 mg · mL-1.Compared with the normal group,the proteins expression levels of double-strand RNA-activated protein kinase-like ER kinase (PERK),glucose-regulated protein78 (GRP78),activating transcription factor-4 (ATF4),C/EBP-homologous protein (CHOP) and apoptosis-related poly-ADP-ribos polymerase (PARP),cleaved cysteinyl aspartate specific proteinase-3 (Caspase-3) were significantly up-regulated after treatment of GPM in vitro (P ＜0.05 or P ＜0.01).PERK and GAPDH grayscale average ratio in normal group,control group,and three concentration experimantal groups were (4.31 ±0.81) ×10-2,(8.92±0.91) ×10-2,(20.73±0.97) ×10-2,(24.04±0.95) ×10-2,(11.65±1.67) × 10-2;GRP78 and GAPDH grayscale average ratio in the five groups were (27.99 ±2.36) × 10-2,(35.58 ± 1.02) × 10-2,(42.55 ± 1.19) × 10-2,(54.91 ± 1.20) × 10-2,(7.31 ± 1.01) × 10-2;ATF4 and GAPDH grayscale average ratio in the five groups were (20.82 ± 1.42) × 10-2,(39.60 ± 0.56) × 10-2,(52.02 ± 1.83) × 10-2,(73.39 ± 1.83) × 10-2,(18.13 ± 2.28) × 10-2;CHOP and GAPDH grayscale average ratio in the five groups were (8.71 ±0.76) × 10-2,(11.27 ± 1.07) × 10-2,(41.29 ± 1.36) × 10-2,(48.55 ± 1.37) × 10-2,(33.01 ±3.95) × 10-2;PARP and GAPDH grayscale average ratio in the five groups were (13.06 ± 2.88) × 10-2,(36.79 ± 2.10) × 10-2,(58.72 ± 1.53) × 10-2,(67.61 ± 1.68) × 10-2,(34.88 ± 2.02) × 10-2;C aspase-3 and GAPDH grayscale average ratio in the five groups were (5.92 ±0.33) × 10-2,(14.71 ±1.11) × 10-2,(17.58±1.33) × 10-2,(35.41 ±2.91) × 10-2,(5.94 ± 1.61) × 10-2.Compared with the normal group,the expression levels of GRP78,ATF4,CHOP in the four groups were significantly up-regulated (P ＜ 0.05 or P ＜ 0.01).Conclusion GPM can inhibit proliferation and induce apoptosis of HepG2 cells,which may be associated with inducing the HepG2 cells endoplasmic reticulum stress.
