21抑制剂联合伊马替尼对慢性粒系白血病K562细胞增殖的作.doc

MicroRNA-21抑制剂联合伊马替尼对慢性粒系白血病K562细胞增殖的作用 赵广富1，朱贵华1，苏林2 （1.连云港市第二人民医院，江苏连云港 222023；2.中国医学科学院北京协和医学院基础医学研究所，北京100005） 摘要： 目的 探讨microRNA-21基因拮抗联合酪氨酸激酶抑制剂伊马替尼对慢性粒细胞白血病细胞株K562细胞的增殖及周期的调控作用。 方法 在293TN细胞中包装并收获可以抑制内源microRNA-21表达的重组慢病毒颗粒，进行病毒滴度测定后，感染K562，经流式分选得到GFP阳性的K562细胞株。Northern blot证实感染后microRNA-21的表达，MTT法检测细胞存活率，流式细胞仪检测细胞周期。 结果 Northern blot证实K562细胞中microRNA-21的表达水平与正常人外周血细胞相比明显升高。抑制microRNA-21表达的K562细胞株生长速度明显减慢，细胞周期运行也受到阻滞；伊马替尼作用于microRNA-21表达抑制的K562细胞后，细胞增殖与伊马替尼单独作用的K562细胞及单独抑制microRNA-21表达的K562细胞相比均明显降低，周期也受到更严重的阻抑。 结论 在K562细胞中联合使用拮抗microRNA-21表达和酪氨酸抑制剂伊马替尼能够协同抑制细胞增殖及细胞周期运行。 关键词：microRNA-21; 伊马替尼; K562; Effect of micorRNA-21 inhibitor combined with imatinib on chronic myeloid leukemia K562 cell growth ZHAO Guang-fu1, ZHU Gui-hua1, SU Lin2 (1.Lianyungang Second People’s Hospital, Lianyungang Jiangsu 222023; 2.Chinese Academy of Medical Sciences (CAMS) Peking Union Medical College (PUMC), Beijing 100005) Abstract： Objective To study the roles of microRNA-21 in cooperation with imatinib in a chronic myeloid leukemia cell line K562. Methods The recombinant lentivirus particles (pMIRZip-miR-21 and control pMIRZip vector) were harvested from 293TN cells and added to the medium with cultured K562 cells. K562 cells expressing GFP were selected by flow cytometry. The expression of microRNA-21 was detectable by Northern blot. The cell proliferation was tested by MTT, and the cell cycle was measured by flow cytometry. Results MicorRNA-21 was significantly up-regulated in K562 cells compared with peripheral blood cells from normal. Inhibition of micorRNA-21 in K562 cells decreased cell proliferation and cell cycle progression. The growth of K562 cells tranduced with Lenti-miR-21 was markedly inhibited after imatinib treatment. Moreover, The percentage of cells at G0/G1 phase greatly increased after transduction with Lenti-miR-21 to K562 combined with imatini. Conclusion The inhibition of micorRNA-21 in combination with imatinib has a synergistic effect on the proliferation an