The miR-125a-3p Inhibits TIM-3 Expression in AML Cell Line HL-60 In Vitro
The miR-125a-3p Inhibits TIM-3 Expression in AML Cell Line HL-60 In Vitro
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DOI：
10.1007/s12288-016-0733-4
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发表时间：
2017-09-01
影响因子：
0.9
通讯作者：
Doosti, Abbas
中科院分区：
文献类型：
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作者：
Emamdoost, Fatemeh;Khanahmad, Hossein;Doosti, Abbas
急性髓系白血病是一种进展迅速的白血病干细胞癌。它的特点是骨髓中未成熟的髓系细胞过度生产，从而排挤正常的造血干细胞(HSC)。TIM-3是一种免疫调节分子，是急性髓系白血病中LSC的特异性表面标志，与HSCs相比，其在AML细胞上的表达较高。研究表明，microRNAs(MiRNAs)可能在癌症进展或抑制中发挥重要作用。基于生物信息学分析，我们预测miR-125a-3p可能是一个对Tim-3表达具有高抑制活性的miRNA。本研究旨在探讨miR-125a-3p对急性髓系白血病细胞株HL-60 TIM-3基因表达的抑制作用。HL-60细胞在含10%胎牛血清的RPMI1640中培养。用佛波酯诱导细胞TIM-3的表达。将miR-125-3p导入细胞24 h，分别用Q-RT-PCR和流式细胞仪检测TIM-3基因和蛋白的表达。本研究结果表明，miR-125a-3p对HL-60细胞TIM-3基因和蛋白表达具有较强的沉默作用。结果表明，miR125a-3p能显著抑制AML细胞TIM-3的表达。因此，我们的结果证实了用Mirwalk和Target Scan软件预测miR-125a-3p对Tim-3的抑制作用的生物信息学预测。
Acute Myeloid Leukemia is a cancer of leukemic stem cells (LSCs) with a rapid progression. It is characterized by overproduction of immature myeloid cells in bone marrow which crowds out normal hematopoietic stem cells (HSC). TIM-3, an immune regulatory molecule, is an LSC specific surface marker in AML with high expression on these cells compared to HSCs. Studies have indicated that micro RNAs (miRNAs) may play an important role in either cancer progression or suppression. Based on bioinformatics assessments, we have predicted that miR-125a-3p could be a miRNA with high suppressive activity on TIM-3 expression. The purpose of this study was to investigate the inhibitory effect of miR-125a-3p on TIM-3 gene expression in an AML cell line, HL-60, in vitro. HL-60 cells were cultured in RPMI 1640 supplied with 10 % FBS. TIM-3 expression was induced on the cells using phorbol miristate acetate. The cells were transfected with miR-125-3p for 24 h and the gene and protein expression of TIM-3 were measured using q-RT-PCR and flow-cytometery methods, respectively. The results of this study showed that miR-125a-3p has a strong silencing effect on TIM-3 gene and protein expression on HL-60 cell line. Based to our results, miR125a-3p can strongly silence TIM-3 expression in AML cell line. Thus, our results have confirmed the bioinformatics prediction of suppressive effect of miR-125a-3p on TIM-3with Mirwalk and Target Scan softwares.