Mechanism of ER stress involved in ATO inhibited HL-60 proliferation
10.3969/j.issn.1001-1978.2017.11.022
- VernacularTitle:内质网应激在三氧化二砷抑制HL-60增殖过程中的效应机制
- Author:
shan Shan ZHANG
1
;
yi Tian LI
;
qin Xiao WANG
;
Bo ZHANG
Author Information
1. 石河子大学药学院
- Keywords:
acute promyelocytic leukemia;
arsenic trioxide;
ER stress;
differentiation;
apoptosis;
MCODE algorithm
- From:
Chinese Pharmacological Bulletin
2017;33(11):1589-1595
- CountryChina
- Language:Chinese
-
Abstract:
Aim To explore the pharmacological mechanism of arsenic trioxide (ATO) on endoplasmic reticulum(ER) stress and provide indirect pharmacological indicators for the therapeutic drugs monitoring of acute promyelocytic leukemia.Methods The ATO toxicology database was utilized to associate gene data and establish protein-protein interaction networks.MCODE algorithm was used to analyze clustering of network topology relationship in order to find the critical functional gene group.The HL-60 cells were used as the model in this study.The cytotoxicity of ATO to HL-60 was evaluated by MTT assay.Real-time PCR assay was involved in detecting the expression of stress-related genes in ER stress.Western blot was used to detect the expression of stress-related proteins in ER stress.ER-specific staining was conducted by ER-Tracker Red.Cell differentiation and apoptosis was tested by flow cytometry.Results 206 ATO-related genes had 3 794 pairs of relationships,and ER stress-related module was outstanding among the top 4 gene cluster.ATO inhibited HL-60 cell proliferation in a dose-dependent manner;4 h after administration ATO could induce ER stress.The expressions of GRP78 and GRP94 were significant and the gene expression level changed greatly over time.The expression level of GRP78,GRP94 and Nrf2 was induced by 1 ～4 μmol · L-1 ATO,and the expression of CHOP was significant by 8 μmol · L-1 ATO.ATO could induce the phosphorylation level of eIF2α protein at 1 ～2 μmol · L-1 as well as a large expression of CHOP protein markedly at 8 μmol · L-1.As shown in ER-specific staining,ATO at 4,8 μmol · L-1 induced ER swelling.Flow cytometry showed that ATO could induce cell differentiation at 1 μmol · L-1,and the differentiation group was obvious at 2 μmol · L-1.So as to the concentration of 4 and 8 μmol · L-1,but the apoptotic group emerged at 8 μmol · L-1.Conclusion ATO can inhibit the proliferation of HL-60 and in this process,ER stress is involved,which shows a concentration-dependent phaseswitching effect.
