摘要
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48h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.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2016年第4期273-277,共5页
Chinese Journal of Hematology
作者简介
通信作者：高子芬，Email：wjshgao@bjmu．edu．cn
