Shafieizadegan Saba, Aberuyi Narges, Rahgozar Soheila
Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Hezar Jarib Street, Isfahan, 81746- 73441, Iran.
Ann Hematol. 2025 Apr;104(4):2417-2427. doi: 10.1007/s00277-024-06181-1. Epub 2025 Feb 5.
MiR-326 downregulation is strongly associated with multidrug resistance (MDR) and has been identified as an adverse prognostic biomarker for pediatric acute lymphoblastic leukemia (pALL). The choice to study miR-326 as a tumor suppressor in cancer biology, particularly its regulation of apoptosis, drug resistance, and stemness, stems from its strong association with MDR and potential as a therapeutic target in pALL. The current study aimed to investigate, for the first time, the molecular mechanisms underlying the role of miR-326 in ALL, using Gene Ontology annotation network and multilayer network analysis. Our findings revealed that miR-326 exhibits a multifunctional anti-tumor behavior, affecting various aspects of drug resistance, stemness, and apoptosis in cancer, particularly in the context of ALL. Quantitative real-time PCR demonstrated downregulation of the ABC transporter mRNAs ABCC1 and ABCB1 but not ABCA3 in B-ALL cells transfected with miR-326 mimic, as confirmed by bioinformatic data. Western blot analysis showed a possible cross talk between miR-326 and P53 through the upregulation of Mdm2 and P53 proteins. The heightened functional activity of P53 was subsequently validated through the observed augmentation in levels of P21 and CCND1, alongside the evident disruption in the expression levels of Bcl-2, Bcl-xl, and Bax genes. Subsequently, the ceRNA network between miR-326 and LncRNAs was exhibited and the impact of exogenous miR-326 on the expression levels of its molecular sponges, H19 and SNHG1 was examined using RT-qPCR. Future studies will explore the potential impact of miR-326 on its targets, and how this may influence the development of novel therapeutic strategies for ALL.
miR-326表达下调与多药耐药（MDR）密切相关，并且已被确定为小儿急性淋巴细胞白血病（pALL）的不良预后生物标志物。在癌症生物学中选择研究miR-326作为肿瘤抑制因子，特别是其对细胞凋亡、耐药性和干性的调节，源于其与MDR的密切关联以及作为pALL治疗靶点的潜力。本研究旨在首次使用基因本体注释网络和多层网络分析，探究miR-326在ALL中作用的分子机制。我们的研究结果表明，miR-326表现出多功能抗肿瘤行为，影响癌症中耐药性、干性和细胞凋亡的各个方面，特别是在ALL的背景下。定量实时PCR显示，用miR-326模拟物转染的B-ALL细胞中ABC转运蛋白mRNA ABCC1和ABCB1下调，但ABCA3未下调，这得到了生物信息学数据的证实。蛋白质免疫印迹分析表明，miR-326与P53之间可能通过Mdm2和P53蛋白的上调存在相互作用。随后，通过观察到的P21和CCND1水平升高以及Bcl-2、Bcl-xl和Bax基因表达水平的明显破坏，验证了P53功能活性的增强。随后，展示了miR-326与长链非编码RNA（LncRNAs）之间的竞争性内源RNA（ceRNA）网络，并使用逆转录定量PCR（RT-qPCR）检测了外源性miR-326对其分子海绵H19和SNHG1表达水平的影响。未来的研究将探索miR-326对其靶点的潜在影响，以及这可能如何影响ALL新型治疗策略的开发。
