MicroRNA-221(miR-221)抑制p27^(Kip1)蛋白促进慢性粒细胞白血病的发生  

Expression of microRNA-221 in chronic myeloid leukemia and its mechanism

作　　者：秦娜[1] 李广波[2] 郭树霞[1] 张晓娟[1] 

机构地区：[1]郑州人民医院血液内科,郑州450003 [2]郑州市儿童医院肾脏风湿科,郑州450003

出　　处：《首都医科大学学报》2017年第4期580-585,共6页Journal of Capital Medical University

摘　　要：目的探讨microRNA-221(miR-221)与慢性粒细胞白血病(chronic myeloid leuklemia,CML)的相关性及其作用机制。方法采用荧光实时定量PCR(quantificational real-time polymerase chain reaction,qRT-PCR)法检测35例CML病人与25例对照组病人骨髓中miR-221的表达情况。K562细胞转染miR-221模拟物和miR-221抑制剂48 h,qRT-PCR法检测miR-221的表达。分别用MTT法检测转染后的K562细胞增生能力,利用流式细胞仪检测转染后K562细胞周期和凋亡。利用数据库预测miR-221的调节基因为p27^(Kip1)。qRT-PCR和Western blotting法检测miR-221对p27^(Kip1)的mRNA和蛋白的表达的影响。结果 CML病人组miR-221表达明显高于对照组。转染miR-221抑制剂组K562细胞凋亡比例显著升高,细胞周期阻滞于G2SM期,细胞增生能力明显受到抑制。而miR-221过表达则明显促进了K562细胞的增生和细胞周期进程。抑制miR-221表达后K562细胞中p27^(Kip1)蛋白表达显著升高,而当miR-221过表达后p27^(Kip1)表达也随之明显降低。结论 miR-221在CML病人的骨髓中高表达,并抑制下游p27^(Kip1)蛋白的表达从而促进白血病K562细胞增生,促进细胞进入细胞周期,可能是CML发生、发展的一个潜在靶点。Objective MicroRNAs （miRNAs, miRs） have emerged as critical regulators of tumor cell proliferation.This paper is to investigate how miR-221 is involved in the process of chronic myeloid leukemia （CML） and its working mechanisms.Methods Quantificational real-time polymerase chain reaction,qRT （qRT-PCR） was used to measure the expression of miR-221 in CML patients and control patients.K562 cells were transfected with miR-221 mimics, inhibitors, or negative controls.MTT assay was used to determine cell viability.Flow cytometry was used to measure the cell apoptosis and cell cycle.Bioinformatics was used to predict the target gene of miR-221, the qRT-PCR and Western blot were used to determine the expression of p27Kip1 expression.Results The expression of miR-221 was increased significantly in the bone marrow cells in CML patients compared with control patients.Overexpression the miR-221 significantly increased cell vitality and promoted cell proliferation and G1-to-S phase transition of the cell cycle in K562 cells, while inhibition of miR-221 rescued the results.p27Kip1 is the important target gene regulated by miR-221, the inhibition of the miR-221 promoted the p27Kip1 expression, while miR-221 overexpression decreased the p27Kip1 expression.Conclusion Our study suggested that miR-221 may be an oncogenic miRNA by inhibiting the protein expression of p27Kip1 in CML.

关 键 词：慢性粒细胞白血病 MIR-221 K562细胞 P27KIP1 

分 类 号：R733.7[医药卫生—肿瘤]