靶向抑制微小RNA-21提高白血病细胞对三氧化二砷的敏感性研究

目的 探讨以微小RNA 21(microRNA 21, miRNA 21)为靶标的反义核酸(AMO miR 21)提高白血病K562细胞对三氧化二砷（As2O3）的敏感性及可能的作用机制。方法 利用LipofectamineTM 2000将化学合成的反义核酸转染K562细胞。MTT法检测单独使用AMO-miR-21、As2O3以及两者联合使用对细胞增殖的抑制作用,并计算抑制率和IC50；PI单染检测细胞周期及凋亡；实时定量PCR检测miRNA-21表达水平的改变；免疫荧光法检测miRNA 21候选靶基因PDCD4蛋白表达的改变。结果 AMO-miR-21与As2O3联合使用后，IC50从2.10 μmol/L降低到1.23 μmol/L，敏感性提高到单用As2O3的1.78倍；AMO-miR-21联合As2O3组细胞凋亡明显增加（P<0.05）；单独转染AMO-miR-21可显著抑制K562细胞中miRNA-21的表达水平（P<0.01)，同时抑癌基因PDCD4蛋白表达明显增加（P<0.05）。结论 AMO-miR-21与As2O3联合使用可提高K562细胞对As2O3的敏感性，为白血病的治疗提供了新的方法。AMO-miR-21通过靶向抑制miRNA-21，进而上调抑癌基因PDCD4的表达而发挥抗肿瘤作用。

Study on the Sensitivity of Leukemic Cells to Arsenic Trioxide Enhanced by Targeted Suppression of miRNA-21

Objective To study the effect of antisense oligonucleotide targeted on miRNA-21 (AMO-miR-21) for enhancing the arsenic trioxide (As2O3) sensitivity of leukemic K562 cells and its possible acting mechanism. Methods Chemosynthetic AMO-miR-21 was transfected to K562 cells using Lipofectamine TM 2000. The inhibitory effects of As2O3 and AMO-miR-21, used singly or in combining, on cell proliferation were detected by MTT, their inhibition rate and IC50 were calculated. Cell cycle and apoptosis were assessed with ...