miR-124-3p靶向ABCA2增强慢性白血病细胞K562-R对伊马替尼的敏感性

目的:探讨miR-124-3p靶向ABCA2对CML细胞株耐药性及细胞活性的影响。方法:过表达miR-124-3p和ABCA2的CML细胞株K562-R及裸鼠皮下移植瘤模型用于本研究。设置K562-R空白对照组、miR-124-3p mimic对照组、ABCA2过表达组和mimic+pc ABCA2组,研究miR-124-3p和ABCA2对K562-R细胞的影响。Western Blot检测体系中增殖、凋亡和自噬相关分子表达;qRT-PCR检测体系中miR-124-3p和ABCA2的表达;荧光素酶系统及生物信息学软件预测miR-124-3p和ABCA2之间靶向关系。CCK-8染色检测细胞增殖,Hoechst染色检测细胞凋亡,免疫组织化学检测细胞凋亡和增殖相关分子。结果:miR-124-3p在K562耐药细胞(K562-R)中低表达,在K562敏感细胞(K562-S)中高表达;ABCA2在K562-R中高表达,在K562-S中低表达(P<0.01);生物信息学和荧光素酶报告系统检测显示,miR-124-3p靶向调节ABCA2表达。miR-124-3p过表达能有效抑制K562-R细胞增殖,促进细胞凋亡和自噬发生(P<0.01);而ABCA2过表达则促进K562-R细胞增殖,明显抑制凋亡和自噬发生(P<0.01);进一步通过裸鼠体内皮下移植瘤模型表明,miR-124-3p可以有效抑制K562-R细胞增殖,促进细胞凋亡和自噬(P<0.01)。结论:miR-124-3p可以通过靶向ABCA2来有效抑制白血病耐药细胞株K562-R的增殖,促进细胞凋亡和自噬。

MiR-124-3p Enhances the Sansitivity of Chronic Myelogenous Leukemia Cell K562-R to Imatinib by Targeting ABCA2

Objective:To investigate the effect and mechanism of miR-124-3p-targeing regulating ABCA2 on chronic myelogenous leukemia cell K562-R.Methods:CML cells with miR-124-3p-overexpression and ABCA2-over-expression as well as subcutaneoustrans planted tumor nude mice were used as study objects.And the CML cells were divided into four groups:K562-R blank control,miR-124-3p mimic control,ABCA2-overexpression and mimic+PC ABCA2.The effects of miR-124-3p and ABCA2 on CML cells were analyzed.The levels of proliferation-,apoptosis-and autophagyrelated protein were determined by Western blot.qRT-PCR was employed to detect the levels of miR-124-3p and ABCA2 in K562-R cells.The relationship between miR-124-3p and ABCA2 was validated by luciferase reporter system assays and bioinformatics.Hoechst/immunohistochemical staining and CCK-8 assay were performed to investigate the function involved.Results:miR-124-3p highly expressed in K562-S cells and lowly expressed in K562-R cells,however,ABCA2 lowly expressed in K562-S cells and highly expressed in K562-R cells.Over-expression of miR-124-3p significantly decreased ABCA2 level and cell growth,but increased autophagy and apoptosis in K562-R cells(P<0.01).When ABCA2 was over-expressed,the K562-R cell growth was promoted and autophagy and apoptosis were inhibited(P<0.01).The miR-124-3p promoted cell autophagy and apoptosis but inhibited cell growth in nude mice transplant tumor model(P<0.01).Conclusion:miR-124-3p can target ABCA2 to inhibit the growth of CML cells and promote the cell autophagy and apoptosis of CML cells.