The induction effect of hydroxyurea and metformin on fetal globin in the K562 cell line.

IF 6 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Mohammad Eini, Hossain Safarpour, Ebrahim Miri-Moghddam
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引用次数: 0

Abstract

Despite the established efficacy of hydroxyurea (HU) in increasing fetal hemoglobin (Hb F) levels in patients with intermedia beta-thalassemia (β-thal) and sickle cell anemia, the precise molecular mechanisms underlying these effects remain largely elusive. Understanding these mechanisms is paramount for identifying alternative therapeutic approaches to increase Hb F production while minimizing adverse effects. In this study, we employed weighted gene co-expression network analysis (WGCNA) to investigate the molecular underpinnings of γ-globin switching within GSE90878 dataset. Leveraging this information, we aimed to predict the transcriptome network and elucidate the mechanism of action of HU and Metformin (Met) on this network comprehensively. Through bioinformatic analysis, we identified IGF2BP1 and GCNT2 as key regulators of the γ-globin switching mechanism. To experimentally validate these findings, we utilized the K562 cell line as an erythroid model. Cells were treated with HU (50, 100, and 150 µM) and Met (50, 100, and 150 µM) for 24, 48, and 72 h. The expression levels of the GCNT2, γ-globin, IGF2BP1, miR-199a/b-5p, miR-451-5p and miR-144-3p were quantified using real-time polymerase chain reaction (qPCR). Our results revealed that treatment with HU (150 µM), Met (100 µM), and combination of HU-Met (150/100 µM) significantly increased IGF2BP1 expression by 6.2, 5.3, and 7.1-fold, respectively, after 24 h treatment. Furthermore, treatment with HU (50 µM), Met (50 µM) and HU/Met (50/50 µM) for 24 h led to a 3.3, 1.2, and 5-fold decrease in GCNT2 gene expression, respectively. Notably, the highest levels of γ-globin expression and Hb F production were observed with HU (100 µM), Met (50 µM), and HU/Met (100/50 µM). This study provides compelling evidence that HU and Met significantly enhance γ-globin expression and Hb F production in the K562 cell line. Our findings suggest that these drugs exert their effects by modulating the expression of IGF2BP1 and GCNT2, thus offering valuable insights into potential therapeutic strategies for disorders characterized by low Hb F levels.

羟基脲和二甲双胍对K562细胞株胎儿珠蛋白的诱导作用。
尽管羟基脲(HU)在提高中β-地中海贫血(β-thal)和镰状细胞性贫血患者的胎儿血红蛋白(Hb F)水平方面已经确立了疗效,但这些作用的确切分子机制在很大程度上仍然是难以捉摸的。了解这些机制对于确定增加血红蛋白F产生的替代治疗方法同时最小化不良反应至关重要。在这项研究中,我们采用加权基因共表达网络分析(WGCNA)来研究GSE90878数据集中γ-珠蛋白转换的分子基础。利用这些信息,我们旨在预测转录组网络,并全面阐明HU和Metformin (Met)在该网络中的作用机制。通过生物信息学分析,我们发现IGF2BP1和GCNT2是γ-珠蛋白转换机制的关键调节因子。为了实验验证这些发现,我们利用K562细胞系作为红系模型。细胞分别用HU(50、100和150µM)和Met(50、100和150µM)处理24、48和72小时。使用实时聚合酶链反应(qPCR)定量检测GCNT2、γ-珠蛋白、IGF2BP1、miR-199a/b-5p、miR-451-5p和miR-144-3p的表达水平。我们的研究结果显示,HU(150µM)、Met(100µM)和HU-Met(150/100µM)联合处理24 h后,IGF2BP1的表达分别显著增加6.2倍、5.3倍和7.1倍。此外,HU(50µM)、Met(50µM)和HU/Met(50/50µM)处理24小时分别导致GCNT2基因表达降低3.3倍、1.2倍和5倍。值得注意的是,在HU(100µM)、Met(50µM)和HU/Met(100/50µM)中,γ-珠蛋白的表达和Hb F的产生水平最高。本研究提供了令人信服的证据,证明HU和Met显著提高K562细胞系γ-珠蛋白的表达和Hb F的产生。我们的研究结果表明,这些药物通过调节IGF2BP1和GCNT2的表达来发挥作用,从而为以低Hb F水平为特征的疾病的潜在治疗策略提供了有价值的见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Molecular Medicine
Molecular Medicine 医学-生化与分子生物学
CiteScore
8.60
自引率
0.00%
发文量
137
审稿时长
1 months
期刊介绍: Molecular Medicine is an open access journal that focuses on publishing recent findings related to disease pathogenesis at the molecular or physiological level. These insights can potentially contribute to the development of specific tools for disease diagnosis, treatment, or prevention. The journal considers manuscripts that present material pertinent to the genetic, molecular, or cellular underpinnings of critical physiological or disease processes. Submissions to Molecular Medicine are expected to elucidate the broader implications of the research findings for human disease and medicine in a manner that is accessible to a wide audience.
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