| 李育敏,谷景义,朱雪娇,陈万群,郭敏,姚君琳,林春燕,费嘉.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[J].,2010,30(2):170-173 |
| 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 |
| Study on the Sensitivity of Leukemic Cells to Arsenic Trioxide Enhanced by Targeted Suppression of miRNA-21 |
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| DOI： |
| 中文关键词: 微小RNA-21 反义寡核苷酸 三氧化二砷 白血病 化疗敏感性 |
| 英文关键词:miRNA-21 antisense oligonucleotide arsenic trioxide leukemia chemosensitivity |
| 基金项目:广东省中医药管理局科研项目(No.2008098);广东省科技计划重点引导项目(No.2006B35502010) |
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| 中文摘要: |
| 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2.10μmol/L降低到1.23μmol/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。 |
| 英文摘要: |
| 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 PI stain; expression of miRNA-21 in cells was detected quantitatively by real-time PCR, and the potential target gene PDCD4 protein expression was detected by immuno-fluoremetry. Results Used in combining with AMO-miR-21, the IC50 of As2O3 could be lowered from 2.1 μmol/L to 1.23 μmol/L, and the sensitivity of cells to As2O3 increased to 1.78-fold; with the amount of apoptotic cells increased significantly. Transfection with AMO-miR-21 alone could down-regulate the expression of miRNA-21 in cells (P<0.01), and up-regulate PDCD4 protein expression level significantly. Conclusions Combined use of AMO-miR-21 and As2O3 could increase the sensitivity of K562 cells to As2O3, which provides a novel potential approach for treatment of leukemia. AMO-miR-21 realizes it anti-tumor action by way of targeted inhibition on miRNA-21, and further up-regulates the expression of anti-tumor gene PDCD4. |
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