三氧化二砷诱导K562/ADM耐药细胞凋亡中活性氧的作用

中国临床药理学与治疗学 ›› 2006, Vol. 11 ›› Issue (12): 1375-1379.

三氧化二砷诱导K562/ADM耐药细胞凋亡中活性氧的作用

张亚莉^1,2, 魏虎来², 郭璐²

¹皖南医学院生化教研室, 芜湖 241001, 安徽;
²兰州大学医学实验中心, 甘肃省新药临床前研究重点实验室, 兰州 730000, 甘肃

收稿日期:2006-06-15 修回日期:2006-08-11 出版日期:2006-12-26 发布日期:2020-11-06
通讯作者: 魏虎来, 男, 研究员, 硕士生导师, 研究方向:细胞分子生物学。Tel:0931-8915391 　E-mail:weihulai@lzu.edu.cn
作者简介:张亚莉, 女, 硕士, 研究方向:生物化学与分子生物学研究。Tel:13675532465 E-mail:slj1226@tom.com
基金资助:
甘肃省科技攻关计划项目(NoGS022-A43-137)

Reactive oxygen species play an important role in arsenic trioxide-induced apoptosis of multi-drug resistant K562/ADM cells

ZHANG Ya-li^1,2, WEI Hu-lai², GUO Lu²

¹Department of Biochemistry, Wannan Medical College, Wuhu 241001, Anhui, China;
²Laboratory Center for Medical Science, Lanzhou University, Key Laboratory of Preclinical Study for New Drugs of Gansu Province, Lanzhou 73000,Gansu, China

Received:2006-06-15 Revised:2006-08-11 Online:2006-12-26 Published:2020-11-06

摘要/Abstract

摘要： 目的探讨三氧化二砷(As₂O₃) 诱导K562/ADM 细胞凋亡中活性氧(ROS) 的作用和其与耐药基因mdr1 及其编码的P-糖蛋白(P-gp) 表达的关系。方法应用噻唑蓝(MTT) 比色法检测K562 ADM 细胞增殖活性;Annexin ⅤPI 染色法检测细胞凋亡;RT-PCR 检测mdr1 基因mRNA 的表达;流式细胞术测定P-gP表达、ROS 活性和细胞内阿霉素(ADM) 含量。结果 As₂O₃显著抑制K562/ADM 细胞的增殖,Annexin Ⅴ/PI 双染检测显示凋亡细胞明显增加;ROS活性明显下降;mdr1 mRNA 和P-gP表达明显降低,P-gP功能受抑, 细胞内ADM 含量显著增高。结论 As₂O₃抑制K562/ADM 耐药细胞的增殖活性和促进其凋亡, 其机制可能为通过降低细胞ROS 水平、抑制mdr1/P-gP的表达和功能, 进而逆转mdr1/P-gp介导的多药耐药和凋亡抑制。

关键词: 三氧化二砷, 多药耐药, 凋亡, 活性氧, mdr1, P-糖蛋白

Abstract: AIM: To discuss the role of the reactive oxygen species (ROS) and relationshiPbetween ROS and expression of mdr1/P-gPduring apoptosis induced by arsenic trioxide (As₂O₃) in multidrug-resistant human leukemia K562/ADM cells.METHODS: The cell proliferating activity was assessed with MTT assay.The cell apoptosis was determined by annexin Ⅴ/PI staining.ROS was labelled by DCFH-DA and examined by flow cytometry.Expression of mdr1 mRNA and P-gPwere detected by RT-PCR and flow cytometry, respectively.The contents of adriamycin (ADM) were detected by flow cytometry.RESULTS: As2O3 inhibited K562/ADM cells growth effectively, and the apoptosis rate of the cells by Annexin Ⅴ/PI staining was obviously increased.During apoptosis induced by 2 to 5 μmol·L^-1As₂O₃, the level of ROS was markedly decreased;the expression of mdr1 mRNA and P-gPwere significantly down-regulated, and the function of P-gPwas restrained so that the content of ADM increased in the cells.CONCLUSION: As₂O₃inhibits the proliferation activity and reverses phenomena of P-gp-mediated multidrug-resistance and apoptosis resistance in drug-resistant K562/ADM cells.The possible mechanism is down-regulation of mdr1/P-gPexpression via declines of ROS activity.

Key words: arsenic trioxide, multi-drug resistance, apoptosis, reactive oxygen species, mdr1, P-gp

中图分类号:

张亚莉, 魏虎来, 郭璐. 三氧化二砷诱导K562/ADM耐药细胞凋亡中活性氧的作用[J]. 中国临床药理学与治疗学, 2006, 11(12): 1375-1379.

ZHANG Ya-li, WEI Hu-lai, GUO Lu. Reactive oxygen species play an important role in arsenic trioxide-induced apoptosis of multi-drug resistant K562/ADM cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2006, 11(12): 1375-1379.

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