微小RNA-19a-3p通过调控转录因子ETS样蛋白3对乳腺癌细胞阿霉素耐药的影响  

Effect of microRNA-19a-3p on adriamycin resistance in breast cancer cells by regulating ETS-like 3 transcription factor

作　　者：伏鹏 李文环[1] 李海 Fu Peng;Li Wenhuan;Li Hai(Department of Thyroid and Breast Surgery,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China)

机构地区：[1]华中科技大学同济医学院附属武汉市中心医院甲乳外科,武汉430014

出　　处：《中华实验外科杂志》2023年第9期1741-1744,共4页Chinese Journal of Experimental Surgery

基　　金：武汉市医学科研项目(WX21D69);湖北省卫健委科研项目(WJ2021M009)。

摘　　要：目的探讨微小RNA(miR)-19a-3p在乳腺癌细胞阿霉素耐药中的作用及其机制。方法以人乳腺癌细胞MDA-MB-231为研究对象,根据转染物的不同,将细胞分为对照组、miR-NC组(转染miR-19a-3p mimics阴性对照miR-NC)、miR-19a-3p mimics组(转染miR-19a-3p mimics)和miR-19a-3p mimics+si-转录因子ETS样蛋白3(ELK3)组[共转染miR-19a-3p mimics和ELK3小干扰RNA(siRNA)]。实时定量反转录聚合酶链反应(RT-qPCR)检测细胞中miR-19a-3p的表达,RT-qPCR和蛋白质印迹法(Western blot)检测细胞中ELK3的mRNA和蛋白表达,Western blot法检测细胞中P-糖蛋白(P-gp,ABCB1)的表达,分别用不同浓度的阿霉素(0、5、10、25、50 nmol/L)处理各组细胞,细胞计数试剂盒(CCK-8)法检测细胞增殖能力,评价各组细胞对阿霉素的药物敏感性。多组间均数比较采用单因素方差分析,组间两两比较采用LSD-t法。结果miR-19a-3p mimics组miR-19a-3p表达(2.33±0.12比0.95±0.06,t=18.20,P<0.01)明显高于miR-NC组,ELK3 mRNA表达(0.62±0.05比0.99±0.10,t=6.01,P<0.01)、ELK3蛋白表达(0.32±0.02比0.56±0.02,t=18.76,P<0.01)、ABCB1蛋白表达(0.33±0.02比0.48±0.02,t=9.23,P<0.01)、半数抑制浓度(IC50)值[(3.74±0.44)nmol/L比(9.01±0.40)nmol/L,t=15.34,P<0.01]明显低于miR-NC组,差异均有统计学意义。miR-19a-3p mimics+si-ELK3组miR-19a-3p表达(3.48±0.38比2.33±0.12,t=5.03,P<0.01)高于miR-19a-3p mimics组,ELK3 mRNA表达(0.32±0.02比0.62±0.05,t=10.69,P<0.01)、ELK3蛋白表达(0.15±0.02比0.32±0.02,t=10.05,P<0.01)、ABCB1蛋白表达(0.24±0.03比0.33±0.02,t=4.61,P<0.01)、IC50值[(1.67±0.24)nmol/L比(3.74±0.44)nmol/L,t=7.16,P<0.01]低于miR-19a-3p mimics组,差异均有统计学意义。结论miR-19a-3p能够抑制乳腺癌细胞阿霉素耐药,其机制可能与miR-19a-3p抑制ELK3的表达有关。Objective To investigate the role of miR-19a-3p in adriamycin resistance in breast cancer cells.Methods Human breast cancer cell MDA-MB-231 was used as the research object.According to the different transfectors,the cells were divided into control group,miR-NC group(transfected with miR-19a-3p mimics negative control miR-NC),miR-19a-3p mimics group(transfected with miR-19a-3p mimics)and miR-19a-3p mimics+si-ETS-like 3 transcription factor(ELK3)group[co-transfected with miR-19a-3p mimics and ELK3 small interfering RNA(siRNA)].The expression of miR-19a-3p was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).The mRNA and protein expression of ELK3 was detected by RT-qPCR and Western blotting respectively,and the expression of P-glycoprotein(P-gp,ABCB1)was detected by Western blotting.The cells in each group were treated with different concentrations of adriamycin(0,5,10,25,50 nmol/L),and the proliferation ability of cells was detected by cell counting Kit-8(CCK-8)method to evaluate the sensitivity of each group to adriamycin.One-way ANOVA was used to compare the mean of multiple groups,and LSD-t method was used for pairwise comparison between groups.Results MiR-19a-3p expression in the miR-19a-3p mimics group[(2.33±0.12)vs.(0.95±0.06),t=18.20,P<0.01],ELK3 mRNA expression[(0.62±0.05)vs.(0.99±0.10),t=6.01,P<0.01],ELK3 protein expression[(0.32±0.02)vs.(0.56±0.02),t=18.76,P<0.01],ABCB1 protein expression[(0.33±0.02)vs.(0.48±0.02),t=9.23,P<0.01]and half maximal inhibitory concentration(IC50)value[(3.74±0.44)nmol/L vs.(9.01±0.40)nmol/L,t=15.34,P<0.01]were lower than those in the miR-NC group with the differences being statistically significant.The miR-19a-3p expression in miR-19a-3p mimics+si-ELK3 group[(3.48±0.38)vs.(2.33±0.12),t=5.03,P<0.01]was higher than that in the miR-19a-3p mimics group,and ELK3 mRNA expression[(0.32±0.02)vs.(0.62±0.05),t=10.69,P<0.01],ELK3 protein expression[(0.15±0.02)vs.(0.32±0.02),t=10.05,P<0.01],ABCB1 protein expression[(0.24±0.03)vs.

关 键 词：乳腺癌 微小RNA 转录因子ETS样蛋白3 耐药 

分 类 号：R737.9[医药卫生—肿瘤]