Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran.
Department of Regenerative Biomedicine, Cell Science Research Center, Royan Institute for Stem Cell Biology & Technology, ACECR, Tehran, Iran.
Asian Pac J Cancer Prev. 2022 Apr 1;23(4):1159-1167. doi: 10.31557/APJCP.2022.23.4.1159.
Growing evidence has demonstrated that microRNAs have a major effect on development of different types of cancer including AML. The overexpression of miR-625 could decrease tumorgenesis of acute myeloid leukemia cell lines through Integrin-linked kinase signaling pathway and reducing the associated oncogenes. The aim of this study is to evaluate the effect of hsa-miR-625 upregulation on apoptosis and proliferation of KG1 cell line via ILK signaling pathway.
The KG-1 cell line was transfected with pLenti-III-premir625-GFP through viral method. Then, expression of miR-625 and genes were analyzed by quantitative PCR. Western blotting was used to evaluate for the protein level. Apoptosis was investigated by flow cytometry. Cell cycle analysis with PI and CCK-8 assay were performed to evaluate proliferation.
KG-1 cells transfected with pLenti-III-pre mir625-GFP construct showed a significant increase in cell apoptosis. Gene expression of ILK and NF-κB were downregulated and AKT, c-fos, Caspase3, cyclin D1, KLF-4, OCT-4 and Nanog were upregulated but no alteration in GSK3 expression profile was observed. Downregulation of NF-κB and upregulation of Caspase 3 and p-β-catenin protein levels were indicated (p<0.05).
MiR-625 can be a promising approach to aid in the treatment of AML. However, further studies are required in this respect.
越来越多的证据表明，microRNAs 对包括 AML 在内的不同类型癌症的发展有重大影响。miR-625 的过表达可以通过整合素连接激酶信号通路降低急性髓系白血病细胞系的肿瘤发生，并减少相关的癌基因。本研究旨在通过 ILK 信号通路评估 hsa-miR-625 上调对 KG1 细胞系凋亡和增殖的影响。
通过病毒方法将 pLenti-III-premir625-GFP 转染到 KG-1 细胞系中。然后，通过定量 PCR 分析 miR-625 和基因的表达。通过 Western blot 评估蛋白水平。通过流式细胞术研究凋亡。通过 PI 和 CCK-8 测定法进行细胞周期分析以评估增殖。
转染 pLenti-III-pre mir625-GFP 构建体的 KG-1 细胞显示出明显的细胞凋亡增加。ILK 和 NF-κB 的基因表达下调，AKT、c-fos、Caspase3、cyclin D1、KLF-4、OCT-4 和 Nanog 上调，但 GSK3 表达谱没有改变。NF-κB 的下调和 Caspase 3 和 p-β-catenin 蛋白水平的上调表明（p<0.05）。
miR-625 可能是治疗 AML 的有前途的方法。然而，在这方面需要进一步的研究。
