Kereka Kangup Steven, Mousavi Seyed Hadi, Alizadeh Shaban, Ghaemmaghami Leila, Fakoorizad Ghasem, Motallebzadeh Khanmiri Jamal
Department of Haematology and Blood Transfusion, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran.
Islamic Azad University, North Tehran Branch, Tehran, Iran.
Int J Hematol Oncol Stem Cell Res. 2024 Oct 1;18(4):358-366. doi: 10.18502/ijhoscr.v18i4.16760.
Up-regulation of the microRNA-625 and abnormal expression of the Sox2 gene have been studied and seen in several tumors. Few reports have also shown the aberrant expression of miR-625 and Sox2 expression in various cancers. Several studies have also confirmed that phosphatidylinositol 3' -kinase /protein kinase B pathways regulate hematological malignancies, including Acute Myeloid Leukemia (AML). Thus, this study aimed to investigate the effects of mir-625 up-regulation on proliferation, apoptosis, and cell cycle by targeting the Sox2 gene via the downstream Akt signaling pathway and cell cycle regulators, such as p21, p27, and cyclin E in the KG-1 cell line. Cells obtained from the KG-1 cell line were cultured and transfected with plasmid DNA (miR-625) and scrambled as the control using the Lonza electroporation system. Flow cytometry was used to evaluate cell cycle, proliferation, and apoptosis. Relative gene expression was validated by qRT-PCR. All data were analyzed using graph pad prism 7.01 and REST 2009. : KG-1 cells transfected with the mir625-GFP construct showed decreased proliferation, increased apoptosis, and induced cell cycle arrest. Low levels of Sox2, p21, cyclin E, and up-regulation of p27 were confirmed and validated by qRT-PCR ( P < 0.05 ). MiR-625 can be a promising approach to aid in the treatment of AML. However, further studies are required in this field.
微小RNA-625的上调以及Sox2基因的异常表达已在多种肿瘤中得到研究和观察。也有少数报道显示miR-625和Sox2在各种癌症中存在异常表达。多项研究还证实，磷脂酰肌醇3'-激酶/蛋白激酶B通路调节血液系统恶性肿瘤，包括急性髓系白血病（AML）。因此，本研究旨在通过下游Akt信号通路和细胞周期调节因子（如p21、p27和细胞周期蛋白E）靶向Sox2基因，研究miR-625上调对KG-1细胞系增殖、凋亡和细胞周期的影响。从KG-1细胞系获得的细胞进行培养，并用质粒DNA（miR-625）转染，同时使用Lonza电穿孔系统转染 scrambled作为对照。采用流式细胞术评估细胞周期、增殖和凋亡。通过qRT-PCR验证相对基因表达。所有数据均使用GraphPad Prism 7.01和REST 2009进行分析。结果显示：转染mir625-GFP构建体的KG-1细胞增殖减少、凋亡增加并诱导细胞周期停滞。通过qRT-PCR证实并验证了Sox2、p21、细胞周期蛋白E水平降低以及p27上调（P<0.05）。MiR-625可能是辅助治疗AML的一种有前景的方法。然而，该领域还需要进一步研究。
