MicroRNA-191在T淋巴母细胞性白血病/淋巴瘤中的表达及其作用机制研究

Abstract

目的 探讨MicroRNA-191 (miR-191)与T淋巴母细胞性白血病/淋巴瘤(T-ALL/LBL)的相关性及其作用机制.方法 采用荧光实时定量PCR(qRT-PCR)法检测20例T-ALL/LBL患者肿瘤组织和20例淋巴结反应性增生(LRH)患者淋巴结组织中miR-191的表达,并分析其与临床预后的关系.构建反义miR-191慢病毒载体(LV-miR-191-KD)和阴性对照载体(LV-NC-GFP)并转染T-ALL细胞系Jurkat细胞,qRT-PCR法检测miR-191的表达水平.分别用CCK-8法和流式细胞术检测下调miR-191后的细胞活性、细胞周期和凋亡.结果 T-ALL/LBL患者组miR-191表达水平明显高于LRH患者组(1.875±0.079对1.000,P=0.001),以miR-191表达量的中位数为界,将T-ALL/LBL患者分为高表达组(10例)与低表达组(10例),高表达组患者3年OS率明显低于低表达组(26％对82％,P=0.021).转染48 h后,LV-miR-191-KD组Jurkat细胞miR-191表达水平(0.578±0.012)较LV-NC-GFP组(1.011±0.053)和未转染对照组(1.000)显著降低(P值分别为0.018和0.021),细胞凋亡比例显著升高(P值均＜0.05),细胞周期阻滞于G0/G1期,并抑制从G1期向S期转化.结论 miR-191在T-ALL/LBL的发生、发展过程中起促进作用,可能作为T-ALL/LBL治疗的潜在靶点.

Objective To evaluate the correlation between MicroRNA-191 (miR-191) and T lymphoblastic leukemia/lymphoma (T-ALL/LBL) to probe its underlying molecular mechanism.Methods The expression of miR-191 was examined by real-time PCR (RT-PCR)in 20 T-ALL/LBL tissue samples and 20 lymphoid reactive hyperplasia (LRH) tissue samples.The correlation between miR-191 and the clinicopathological feature of T-ALL/LBL was analyzed.Antisense miR-191 lentiviral vectors was constructed and transfected into T-ALL/LBL Jukat cells.After transfection,the expression of miR-191 was examined by RT-PCR.The cell activity was evaluated by CCK-8 asssy.The cell cycle and apoptosis were determined by flow cytometry.Results Compared with LRH samples,the results of RT-PCR showed significant upregulation of miR-191 in 20 T-ALL/LBL tissue samples (1.875±0.079 vs 1.000,P＜0.05).The expression level of miR-191 was negatively associated with prognosis.Compared with LV-NC-GFP and control groups,the expression of miR-191 significantly decreased after transfection of antisense miR-191 lentiviral vectors (0.578±0.012 vs 1.011±0.053 and 1.000,P＜0.05),the percentages of apoptotic cells and the cell in G0/G1 phase significantly increased(P＜0.05).Conclusions miR-191 might play a significant role in the development of T-ALL/LBL,implicating a new target for therapy.