上调miR-21-5p对阿霉素诱导的心肌细胞增殖、凋亡的影响及其机制研究
Effect and mechanism of up-regulation of miR-21-5p on doxorubicin-induced cardiomyo-cyte proliferation and apoptosis
耿洁 1刘洁婕 2刘亚 3贺赛4
作者信息
- 1. 西安交通大学第二附属医院心内科,西安 710004
- 2. 延安大学附属医院手术室,延安 716000
- 3. 西安交通大学第一附属医院呼吸科,西安 710061
- 4. 陕西省肿瘤医院乳腺病院,西安 710061
- 折叠
摘要
目的:探讨上调miR-21-5p表达对阿霉素(DOX)诱导的心肌细胞增殖和凋亡的影响及其机制研究.方法:将心肌细胞H9C2分成control组(常规培养的对照细胞)、DOX组(用含1 μmol/L DOX的细胞培养液培养)、NC agomir处理组(DOX+NC ago组)、miR-21-5p agomir处理组(DOX+miR-21-5p ago组)、NC antagomir处理组(DOX+NC anta组)、miR-21-5p antagomir处理组(DOX+miR-21-5p anta组).实时荧光定量PCR法测定miR-21-5p的表达,MTT法测定细胞增殖,流式细胞术测定细胞凋亡,western blotting测定细胞中C-Caspase-3、β-catenin、c-Myc蛋白表达.结果:与control组相比,DOX组中miR-21-5p水平降低,细胞增殖活性降低,细胞凋亡和C-Caspase-3蛋白表达水平升高,β-catenin、c-Myc蛋白表达水平下降(P＜0.05).与DOX+NC ago组比较,DOX+miR-21-5p ago组中miR-21-5p水平升高,细胞增殖活性上升,细胞凋亡和C-Caspase-3蛋白表达水平降低,β-catenin、c-Myc蛋白表达水平上升(P＜0.05).结论:上调miR-21-5p可促进DOX诱导的心肌细胞增殖,并通过Wnt/β-catenin信号通路抑制DOX诱导的心肌细胞凋亡,从而改善DOX诱导的心肌毒性作用.
Abstract
Objective:To investigate the effect and mechanism of up-regulation of miR-21-5p on doxorubicin(DOX)-induced cardiomyocyte proliferation and apoptosis.Methods:Cardiomyocytes H9C2 were divided into control group(conventional-cultured control cells),DOX group(cultured with 1 μM DOX),NC agomir group(DOX+NC ago group),miR-21-5p agomir group(DOX+miR-21-5p ago group),NC antagomir treatment group(DOX+NC anta group),and miR-21-5p antagomir treatment group(DOX+miR-21-5p anta group).The expres-sion of miR-21-5p was detected by reverse transcription-quantitative polymerase chain reaction(RT-qPCR),cell proliferation was detected by MTT,cell apoptosis was detected by flow cytometry,and the protein expression of C-Caspase-3,β-catenin and c-Myc was detected by western blotting.Results:Compared with the control group,the level of miR-21-5p and cell proliferation activity were decreased,apoptosis and C-Caspase-3 protein expres-sion levels were increased,and β-catenin and c-Myc protein expression levels were decreased in the DOX group(P＜0.05).Compared with the DOX+NC ago group,the level of miR-21-5p in the DOX+miR-21-5p ago group was increased,cell proliferation activity was increased,apoptosis and C-Caspase-3 protein expression levels were decreased,and β-catenin and c-Myc protein expression levels were increased(P＜0.05).Conclusion:Up-regula-tion of miR-21-5p promotes DOX-induced cardiomyocyte proliferation and inhibits DOX-induced cardiomyo-cyte apoptosis through Wnt/β-catenin signaling pathway,thereby ameliorating DOX-induced cardiotoxicity.
关键词
心肌细胞/miR-21-5p/Wnt/β-catenin/凋亡/阿霉素Key words
cardiomyocyte/miR-21-5p/Wnt/β-catenin/apoptosis/doxorubicin引用本文复制引用
基金项目
陕西省重点研发计划资助项目(2021SF-218)
陕西省自然科学基础研究计划资助项目(2018JQ8044)
出版年
2024
国家科技期刊平台
NSTL
万方数据