摘要
目的通过检测人白血病细胞株K562及阿霉素耐药细胞株K562/A02中miR-155和Ets-1表达,探讨miR-155、Ets-1表达与白血病化疗耐药的关系。方法利用Lipofectamine 2000将含有miR-155模拟物和miR-155抑制物的寡核苷酸探针、Ets-1过表达质粒以及Ets-1干扰质粒转染K562及K562/A02细胞,实时荧光定量PCR法检测miR-155、Ets-1及多药耐药基因(MDR1)的表达,Western blotting检测Ets-1和MDR1蛋白表达水平;转染后的K562及K562/A02细胞经阿霉素处理24 h,MTT法检测细胞存活率。结果与K562细胞相比,K562/A02细胞中miR-155、Ets-1和MDR1表达水平显著升高(P<0.05)。抑制miR-155表达能显著增强阿霉素对K562/A02细胞的抑制作用,并抑制Ets-1和MDR1表达,而MDR1表达抑制效应可被Ets-1过表达所逆转。结论 miR-155通过调节Ets-1表达参与白血病耐药形成,miR-155有可能成为逆转白血病耐药的作用靶点。
Objective To detect the expression of miR-155 and Ets-1 in leukemia cell line K562 and adriamycin resistance K562 / A02 cell line,and explore the relationship between miR-155,Ets-1 and drug resistance in leukemia treatment. Methods miR-155 simulation oligonucleotide probes and miR-155 inhibitor oligonucleotide probes,Ets-1 expression plasmid and Ets-1 interference expression plasmid were transfected in K562 and K562 / A02 cells by Lipofectamine 2000. Real-time quantitative PCR was used to determine the expression of miR-155,Ets-1 and multidrug resistance（ MDR1）. The protein levels of Ets-1 and MDR1 were analyzed by Western blotting. The viability of K562 and K562 / A02 cells was evaluated by MTT assay. Results The expression levels of miR-155,Ets-1 and MDR1 were significantly increased in K562 / A02 cells than those in K562 cells. Down-regulation of miR-155 could obviously increase the inhibition rate of adriamycin on K562 / A02 cells and inhibit the expression of Ets-1 and MDR1,which could be reversed by overexpression of Ets-1. While,up-regulation of miR-155 could significantly increase the inhibition rate of adriamycin on K562 cells and stimulate the expression of Ets-1 and MDR1,which could be reversed by Ets-1 siRNA. Conclusion miR-155 is involved in the mechanism of drug resistance in leukemia by regulating Ets-1 expression,which may provide a potential novel target for overcoming drug-resistance.
出处
《临床肿瘤学杂志》
CAS
2015年第12期1063-1067,共5页
Chinese Clinical Oncology
