Moghaddam Yaser, Andalib Alireza, Mohammad-Ganji Maryam, Homayouni Vida, Sharifi Mohammadreza, Ganjalikhani-Hakemi Mazdak
Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Acqured Immunodeficiency Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.
Pathol Res Pract. 2018 Sep;214(9):1482-1488. doi: 10.1016/j.prp.2018.07.019. Epub 2018 Jul 24.
Acute Myeloid Leukemia (AML) is a Cancer of hematopoietic stem cells with a rapid progression. TIM-3 is expressed on leukemic stem cells (LSCs) in most types of AML and might have a positive effect on maintenance of malignant phenotype. MicroRNAs play important roles in either cancer progression or suppression. In this study were evaluated, the inhibitory effect of miR-498 on TIM-3 expression and its impact on proliferation and survival of HL-60 cell line.
Firstly, the probable inhibitory effect of miR-498 on TIM-3 expression was predicted. HL-60 cells were cultured and expression of TIM-3 was induced on the cells using phorbol miristate acetate. The cells were transfected with miR-498 and expression level of TIM-3 were measured using with q-RT-PCR and flow cytometry methods. In addition, the effect of suppression of TIM-3 expression in HL-60 cell line was analyzed with apoptosis and cell proliferation assays.
Bioinformatics analyses predicted that miR-498 has high ability to silence TIM-3 gene expression. Our experiments confirmed that miR-498 was able to strongly silence TIM-3 expression (68% silencing) in HL-60 cell line (P < 0.002). Also, the cells with suppressed expression of TIM-3 had a lower proliferation and higher apoptosis rates.
Based on our results, the miR-498 can effectively suppress TIM-3 expression in the AML cell line. TIM-3 suppression, in turn, inhibits malignant cell proliferation and induces its apoptosis. Collectively, suppression of TIM-3 by miR-498 can be considered as a potential powerful way for treatment of AML.
急性髓系白血病（AML）是一种造血干细胞癌症，进展迅速。TIM-3在大多数类型的AML的白血病干细胞（LSC）上表达，可能对维持恶性表型有积极作用。微小RNA在癌症进展或抑制中起重要作用。在本研究中，评估了miR-498对TIM-3表达的抑制作用及其对HL-60细胞系增殖和存活的影响。
首先，预测miR-498对TIM-3表达的可能抑制作用。培养HL-60细胞，并用佛波酯诱导细胞上TIM-3的表达。用miR-498转染细胞，并用q-RT-PCR和流式细胞术方法测量TIM-3的表达水平。此外，通过凋亡和细胞增殖试验分析HL-60细胞系中TIM-3表达抑制的效果。
生物信息学分析预测miR-498具有高度沉默TIM-3基因表达的能力。我们的实验证实，miR-498能够在HL-60细胞系中强烈沉默TIM-3表达（沉默68%）（P < 0.002）。此外，TIM-3表达受抑制的细胞增殖率较低，凋亡率较高。
根据我们的结果，miR-498可以有效抑制AML细胞系中TIM-3的表达。TIM-3的抑制反过来又抑制恶性细胞增殖并诱导其凋亡。总的来说，miR-498抑制TIM-3可被认为是治疗AML的一种潜在有效方法。
