miR-1306-5p在AML患者骨髓组织单个核细胞中的表达及对HL60细胞增殖凋亡和迁移侵袭调控作用观察

Expression of miR-1306-5p in mononuclear cells of bone marrow tissues of AML patients and its regulation on proliferation,apoptosis,migration,and invasion of HL60 cells

目的观察微小RNA-1306-5p(miR-1306-5p)在急性髓系白血病(AML)患者骨髓组织单个核细胞中的表达及对人白血病细胞株HL60增殖凋亡和迁移侵袭调控作用。方法分别提取初诊AML患者和再生障碍性贫血(AA)患者骨髓组织单个核细胞,采用qRT-PCR法检测单个核细胞miR-1306-5p;(2)取HL60细胞,分别转染过表达和敲低miR-1306-5p质粒(mimics control、miR-1306-5p mimics、inhibitor control、miR-1306-5p inhibitor,计为1、2、3、4组),转染48 h后采用CCK8法检测各组细胞OD值,流式细胞术和Transwell法分别检测各组细胞凋亡率、迁移细胞数、侵袭细胞数。结果AML患者和AA患者miR-1306-5p相对表达量分别为0.27±0.07、1.00±0.08,两者比较,P<0.05。与mimics control组比较,miR-1306-5p mimics组OD值、侵袭细胞数、迁移细胞数降低,凋亡率升高(P均<0.05);与inhibitor control组比较,miR-1306-5p inhibitor组OD值、侵袭细胞数、迁移细胞数升高,凋亡率降低(P均<0.05)。结论AML患者骨髓组织单个核细胞miR-1306-5p表达降低,转染过表达miR-1306-5p质粒的HL60细胞增殖、迁移、侵袭能力减弱及凋亡能力增强,敲低miR-1306-5p质粒的HL60细胞增殖、迁移、侵袭能力增强及凋亡能力减弱。

Objective To observe the expression of microRNA-1306-5p in mononuclear cells of bone marrow tissues in patients with acute myeloid leukemia(AML)and its regulatory effect on proliferation,apoptosis,migration,and invasion of human leukemia cell line HL60.Methods①Bone marrow mononuclear cells from patients with newly diagnosed AML and from patients with aplastic anemia(AA)were extracted,and miR-1306-5p was detected by realtime PCR.②The human leukemic cell line HL60 was divided into the groups 1,2,3 and 4,which were transfected with the mimics control,miR-1306-5p mimics,inhibitor control,and miR-1306-5p inhibitor,respectively;the OD value of each group was determined by CCK8 at 48 h after transfection.Flow cytometry and Transwell assay were used to determine the apoptosis and the number of invasive cells and migratory cells.Results The relative expression levels of miR-1306-5p in the AML patients and AA patients were 0.27±0.07 and 1.00±0.08,respectively,with statistically significant difference(P<0.05).Compared with the mimics control group,the OD value,the number of invasive cells and migratory cells of the miR-1306-5p mimics group decreased and the apoptosis rate increased(all P<0.05).Compared with the inhibitor control group,the OD value,the number of invasive cells and migratory cells of the miR-1306-5p inhibitor group increased,and the apoptosis rate decreased(all P<0.05).Conclusion The expression of miR-1306-5p in mononuclear cells of the bone marrow tissues of AML patients is reduced,the proliferation,migration,invasion ability and apoptosis ability of HL60 cells transfected with overexpression of miR-1306-5p plasmid decrease,and the proliferation,migration,invasion ability,and apoptosis ability of HL60 cells with knockdown of miR-1306-5p plasmid are enhanced.