Arjmand Fateme, Shojaei Samaneh, Khalili Mitra, Dinmohammadi Hossein, Poopak Behzad, Mohammadi-Yeganeh Samira, Mortazavi Yousef
Department of Medical Genetics and Molecular medicine, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Bioimpacts. 2024;14(4):28870. doi: 10.34172/bi.2023.28870. Epub 2023 Nov 28.
The PI3K/AKT/mTOR signaling pathway plays a significant role in the development of T-cell acute lymphoblastic leukemia (T-ALL). Rapamycin is a potential therapeutic strategy for hematological malignancies due to its ability to suppress mTOR activity. Additionally, microRNAs (miRNAs) have emerged as key regulators in T-ALL pathophysiology and treatment. This study aimed to investigate the combined effects of rapamycin and miRNAs in inhibiting the PI3K/AKT/mTOR pathway in T-ALL cells.
Bioinformatic algorithms were used to find miRNAs that inhibit the PI3K/AKT/mTOR pathway. Twenty-five bone marrow samples were collected from T-ALL patients, alongside five control bone marrow samples from non-leukemia patients. The Jurkat cell line was chosen as a representative model for T-ALL. Gene and miRNA expression levels were assessed using quantitative real-time PCR (qRT-PCR). Two miRNAs exhibiting down-regulation in both clinical samples and Jurkat cells were transfected to the Jurkat cell line to investigate their impact on target gene expression. Furthermore, in order to evaluate the potential of combination therapy involving miRNAs and rapamycin, apoptosis and cell cycle assays were carried out.
Six miRNAs (miR-3143, miR-3182, miR-99a/100, miR-155, miR-576-5p, and miR-501- 3p) were predicted as inhibitors of PI3K/AKT/mTOR pathway. The expression analysis of both clinical samples and the Jurkat cell line revealed a simultaneous downregulation of miR-3143 and miR-3182. Transfection investigation demonstrated that the exogenous overexpression of miR-3143 and miR-3182 can effectively inhibit PI3K/AKT/mTOR signaling in the Jurkat cell line. Moreover, when used as a dual inhibitor along with rapamycin, miR-3143 and miR-3182 significantly increased apoptosis and caused cell cycle arrest in the Jurkat cell line.
These preliminary results highlight the potential for improving T-ALL treatment through multi-targeted therapeutic strategies involving rapamycin and miR-3143/miR-3182.
PI3K/AKT/mTOR信号通路在T细胞急性淋巴细胞白血病（T-ALL）的发展中起重要作用。雷帕霉素因其抑制mTOR活性的能力，是血液系统恶性肿瘤的一种潜在治疗策略。此外，微小RNA（miRNA）已成为T-ALL病理生理学和治疗中的关键调节因子。本研究旨在探讨雷帕霉素和miRNA联合抑制T-ALL细胞中PI3K/AKT/mTOR通路的作用。
使用生物信息学算法寻找抑制PI3K/AKT/mTOR通路的miRNA。从T-ALL患者中收集25份骨髓样本，以及来自非白血病患者的5份对照骨髓样本。选择Jurkat细胞系作为T-ALL的代表性模型。使用定量实时PCR（qRT-PCR）评估基因和miRNA表达水平。将在临床样本和Jurkat细胞中均表现出下调的两种miRNA转染到Jurkat细胞系中，以研究它们对靶基因表达的影响。此外，为了评估miRNA与雷帕霉素联合治疗的潜力，进行了凋亡和细胞周期检测。
六种miRNA（miR-3143、miR-3182、miR-99a/100、miR-155、miR-576-5p和miR-501-3p）被预测为PI3K/AKT/mTOR通路的抑制剂。临床样本和Jurkat细胞系的表达分析显示miR-3143和miR-3182同时下调。转染研究表明，miR-3143和miR-3182的外源性过表达可有效抑制Jurkat细胞系中的PI3K/AKT/mTOR信号传导。此外，当与雷帕霉素一起用作双重抑制剂时，miR-3143和miR-3182显著增加Jurkat细胞系中的细胞凋亡并导致细胞周期停滞。
这些初步结果突出了通过涉及雷帕霉素和miR-3143/miR-3182的多靶点治疗策略改善T-ALL治疗的潜力。
