[1]张闪闪,李天一,王晓琴,等.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[J].中国药理学通报,2017,(11):1589-1595.[doi:10.3969/j.issn.1001-1978.2017.11.022]
 ZHANG Shan-shan,LI Tian-yi,WANG Xiao-qin,et al.Mechanism of ER stress involved in ATO inhibited HL-60 proliferation[J].Chinese Pharmacological Bulletin,2017,(11):1589-1595.[doi:10.3969/j.issn.1001-1978.2017.11.022]
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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()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2017年11期
页码:
1589-1595
栏目:
论著
出版日期:
2017-10-20

文章信息/Info

Title:
Mechanism of ER stress involved in ATO inhibited HL-60 proliferation
文章编号:
1001-1978(2017)11-1589-07
作者:
张闪闪1李天一1王晓琴1张 波12
1.石河子大学药学院,2.新疆植物药资源利用教育部重点实验室,新疆 石河子 832002
Author(s):
ZHANG Shan-shan1 LI Tian-yi1 WANG Xiao-qin1 ZHANG Bo12
1.School of Pharmaceutics, Shihezi University;
2.Key Lab of Xinjiang Plant Resources and Utilization, Ministry of Education, Shihezi University, Shihezi Xinjiang 832002,China
关键词:
急性早幼粒细胞白血病 三氧化二砷 内质网应激 分化 凋亡 MCODE算法
Keywords:
acute promyelocytic leukemia arsenic trioxide ER stress differentiation apoptosis MCODE algorithm
分类号:
R329.24;R733.710.22;R979.1
DOI:
10.3969/j.issn.1001-1978.2017.11.022
文献标志码:
A
摘要:
目的 探究三氧化二砷(ATO)引起内质网应激的效应机制,为掌控急性早幼粒白血病治疗药物监测提供间接药理学指标。方法 通过ATO的毒理学数据库关联基因数据整理建立蛋白-蛋白互作网络,使用MCODE算法对网络拓扑关系聚类分析寻找关联基因群; 采用HL-60细胞模型进行实验研究。MTT法评价ATO对HL-60的细胞毒活性; Real-time PCR法检测细胞内质网应激相关基因表达变化; Western blot检测内质网应激相关蛋白表达水平; 用ER-Tracker Red对内质网进行特异性荧光染色; 流式细胞技术检测细胞分化及凋亡指标变化。结果 ATO相关的206个基因PPI网络中有3 794对关系,排名前4的基因群中内质网应激模块凸显; ATO能抑制HL-60细胞增殖,并呈浓度依赖性; ATO在给药4 h即可引起细胞内质网应激,GRP78和GRP94表达较明显,随着时间延长基因表达水平变化明显,1~4 μmol·L-1引起GRP78、GRP94和Nrf2表达,8 μmol·L-1引起CHOP大量表达。ATO在1、2 μmol·L-1时引起eIF2α蛋白磷酸化水平增加,在8 μmol·L-1时诱导CHOP蛋白大量表达; 内质网染色结果显示,4、8 μmol·L-1的ATO诱导内质网的肿胀; 流式结果表明ATO在1 μmol·L-1时引起细胞分化,在2 μmol·L-1时出现明显分化群,4、8 μmol·L-1时细胞分化同样明显,但在8 μmol·L-1时细胞同时出现凋亡群。结论 ATO抑制HL-60细胞增殖过程中出现内质网应激,并呈浓度依赖性阶段性切换效应。
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 phase-switching effect.

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备注/Memo

备注/Memo:
收稿日期:2017-07-18,修回日期:2017-08-22
基金项目:国家自然科学基金资助项目(No 81460566,U1603122)
作者简介:张闪闪(1992-),女,硕士生,研究方向:系统药理学与分子药理学,E-mail:1520813826@qq.com;
张 波(1978-),男,博士,教授,研究方向:系统药理学与中药药理学,通讯作者,E-mail:bozhang_lzu@126.com
更新日期/Last Update: 2017-10-20