Up-Regulation of miR-625-5p Correlates with Suppressed Sox2, Increased Apoptosis, and Cell Cycle Arrest via The PI3K/AKT Signalling Pathway in Acute Myeloid Leukaemia.

Kangup Steven Kereka, Seyed Hadi Mousavi, Shaban Alizadeh, Leila Ghaemmaghami, Ghasem Fakoorizad, Jamal Motallebzadeh Khanmiri
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Abstract

Background: 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. Materials and Methods: 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. Results: 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 ). Conclusion: MiR-625 can be a promising approach to aid in the treatment of AML. However, further studies are required in this field.

在急性髓性白血病中,miR-625-5p上调通过PI3K/AKT信号通路与Sox2抑制、细胞凋亡增加和细胞周期阻滞相关
背景:在一些肿瘤中已经研究并发现了microRNA-625的上调和Sox2基因的异常表达。少数报道也显示了miR-625和Sox2在各种癌症中的异常表达。一些研究也证实了磷脂酰肌醇3' -激酶/蛋白激酶B通路调节血液系统恶性肿瘤,包括急性髓性白血病(AML)。因此,本研究旨在研究mir-625上调对KG-1细胞系增殖、凋亡和细胞周期的影响,通过下游Akt信号通路和细胞周期调节因子p21、p27、cyclin E靶向Sox2基因。材料和方法:培养KG-1细胞系的细胞,转染质粒DNA (miR-625),并用Lonza电穿孔系统炒匀作为对照。流式细胞术检测细胞周期、增殖和凋亡情况。通过qRT-PCR验证相关基因的表达。所有数据采用graphpad prism 7.01和rest2009进行分析。结果:转染mir625-GFP构建物的KG-1细胞增殖减少,凋亡增加,细胞周期阻滞。qRT-PCR证实Sox2、p21、cyclin E低表达,p27上调(P < 0.05)。结论:MiR-625可能是一种有希望的辅助治疗AML的方法。然而,这一领域还需要进一步的研究。
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