miR-155-5p对急性髓系白血病细胞凋亡的影响及可能机制

Effect of miR-155-5p on the apoptosis of acute myeloid leukemia cells and its possible mechanism

目的:探讨miR-155-5p对急性髓系白血病(acute myeloid leukemia,AML)细胞凋亡的影响及可能机制。方法:收集43例AML患者和21例缺铁性贫血患者的骨髓单个核细胞(bone marrow mononuclear cells,BMMNC),qRT-PCR法检测两组细胞miR-155-5p和Bcl-2 mRNA表达水平。AML细胞转染miR-155-5p类似物或抑制剂后,CCK-8法检测经不同浓度阿糖胞苷作用后AML细胞的活性,流式细胞术检测经阿糖胞苷(0.16μg/mL)作用后AML细胞的凋亡情况。通过qRT-PCR和蛋白质印迹法分别检测AML细胞转染miR-155-5p抑制剂后NF-κB、Bcl-2的mRNA和蛋白表达水平。结果:AML患者较缺铁性贫血患者BMMNC的miR-155-5p(t=4.688,P=0.002)和Bcl-2 mRNA(t=4.066,P=0.014)的表达水平明显升高。CCK-8和流式细胞术检测结果显示,转染miR-155-5p类似物的AML细胞经阿糖胞苷处理后细胞活性增高,凋亡明显减少;相反,转染miR-155-5p抑制剂组细胞活性降低,凋亡明显增多(P均<0.01)。qRT-PCR和蛋白质印迹结果显示,转染miR-155-5p抑制剂后AML细胞NF-κB、Bcl-2表达水平降低。结论:miR-155-5p可能通过上调NF-κB、Bcl-2的表达抑制AML细胞凋亡。

Objective:To investigate the effect of miR-155-5p on the apoptosis of acute myeloid leukemia(AML)cells and its possible mechanism.Methods:Bone marrow mononuclear cells of 43 AML patients and 21 iron deficiency anemia patients were collected,and the expression of miR-155-5p and Bcl-2 were detected by quantitative reverse transcriptase PCR(qRT-PCR).After AML cells were transfected with miR-155-5p mimics or inhibitors,CCK-8 assay was used to detect the cell activity of AML cells treated with different concentration of cytarabine,and flow cytometry was used to detect the apoptosis rate of AML cells that treated with cytarabine(0.16μg/mL).The mRNA and protein expression levels of NF-κB and Bcl-2 in AML cells which were transfected with miR-155-5p inhibitor were detected by qRT-PCR and Western blotting,respectively.Results:The mRNA expression level of miR-155-5p and Bcl-2 in AML patients was significantly higher than those in anemia group.Results of CCK-8 and flow cytometry showed that AML cells transfected with miR-155-5p mimic had higher cell activity and lower apoptosis rate after treatment with cytarabine;In contrast,the cell transfected with miR-155-5p inhibitor had lower activity and higher apoptosis rate(P<0.01).The results of qRT-PCR and Western blotting showed that the expression levels of NF-κB and Bcl-2 in miR-155-5p inhibitor group were decreased.Conclusion:miR-155-5p may inhibit AML cell apoptosis by up-regulating the expression of NF-κB and Bcl-2.