Upregulation of hsa-miR-625-5p Inhibits Invasion of Acute Myeloid Leukemia Cancer Cells through ILK/AKT Pathway

Sahar Samieyan Dehkordi, S. Mousavi, Marzieh Ebrahimi, S. Alizadeh, Amir Abbas Hedayati Asl, Monireh Mohammad, Bahareh Aliabedi
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Abstract

Objective Acute myeloid leukemia (AML) is characterized by abnormalities of differentiation and growth of primary hematopoietic stem cells (HSCs) in the blood and bone marrow. In many studies, miR-625-5p has been shown to inhibit downstream pathways from affecting the metastasis and invasion of the integrin-linked kinase (ILK) signaling pathway. It has been proved that the expression of miR-625-5p decreases in AML cell lines. This study aimed to investigate the effect of miR-625-5p upregulation on the invasion of KG1 ell line in vitro. Materials and Methods In this experimental study, we investigated the impact of upregulation of miR-625-5p on invasion via the ILK/AKT pathway in the KG1 cell line. After transfection using the viral method, the cellular invasion was assessed by invasion assay and the levels of miR-625-5p genes and protein were evaluated by quantitative polymerase chain reaction (qPCR) and western blotting. Moreover, CXCR4 level was assessed by flow cytometry. Results The invasion significantly reduced in MiR-625-5p-transfected KG1 cells (P<0.01) that was concomitant with remarkably decreasing in the expression levels of ILK, NF-κB, and COX2 genes compare with the control group (P<0.01). Incontrast, MMP9, AP1, and AKT significantly increased (P<0.01, P<0.001 and P<0.01, respectively) and GSK3β did not change significantly in MiR-625-5p-transfected KG1 cells. The protein level of NF-κB decreased (P<0.01) and MMP9 increased, however it was not significant. Moreoever, the expression of CXCR4 was significantly lower (P<0.01) in comparison with the control group. Conclusion miR-625-5p leads to a reduction in cell invasion in the AML cell line through ILK pathway. Therefore, it could be a breakthrough in future AML-related research. However, further studies are needed to support this argument.
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上调hsa-miR-625-5p通过ILK/AKT通路抑制急性髓系白血病细胞的侵袭
目的急性髓性白血病(Acute myeloid leukemia, AML)以造血干细胞(primary hematopoietic stem cells, hsc)在血液和骨髓中的分化和生长异常为特征。在许多研究中,miR-625-5p已被证明可以抑制下游通路影响整合素连接激酶(ILK)信号通路的转移和侵袭。已经证明,miR-625-5p在AML细胞系中表达降低。本研究旨在探讨miR-625-5p上调对体外KG1细胞系侵袭的影响。材料和方法在本实验研究中,我们通过ILK/AKT通路研究了miR-625-5p上调对KG1细胞系侵袭的影响。采用病毒法转染后,采用侵袭试验评估细胞侵袭,采用定量聚合酶链反应(qPCR)和western blotting检测miR-625-5p基因和蛋白水平。流式细胞术检测CXCR4水平。结果mir -625-5p转染的KG1细胞侵袭性明显降低(P<0.01), ILK、NF-κB、COX2基因表达水平明显低于对照组(P<0.01)。相比之下,转染mir -625-5p的KG1细胞中MMP9、AP1和AKT显著升高(分别为P<0.01、P<0.001和P<0.01), GSK3β无显著变化。NF-κB蛋白水平降低(P<0.01), MMP9蛋白水平升高,但差异不显著。与对照组相比,CXCR4的表达显著降低(P<0.01)。结论miR-625-5p通过ILK途径导致AML细胞系细胞侵袭减少。因此,这可能是未来aml相关研究的一个突破。然而,需要进一步的研究来支持这一论点。
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