摘要
目的探讨伊马替尼在体外对c—kit表达阳性多发性骨髓瘤细胞的作用及机制。方法不同浓度伊马替尼处理KM3细胞后，采用二甲氧唑黄比色法（XTT法）检测药物的细胞毒性作用，流式细胞术检测细胞周期，Annexin V／PI双标流式细胞术和DNA片段分析法检测细胞凋亡，Western blot法检测蛋白质水平变化。结果伊马替尼浓度在0．25μmol／L或以上时，可明显抑制KM3细胞增殖，呈量效关系，48hIC50为0．33μmol／L（P〈0．01）；阻滞细胞周期于G0／G1期；Annexin V和DNA凝胶电泳检测提示随着伊马替尼浓度的增大，KM3细胞凋亡率增加，且可以促使caspase-3和聚ADP核糖聚合酶（PARP）发生裂解；处理后c-kit表达降低，且阻断外源性IL-6对c—kit的促磷酸化作用。结论伊马替尼可以通过抑制c-kit受体相关信号传导途径抑制KM3细胞增殖，诱导细胞凋亡。
Objective To explore the influence of Imatinib on multiple myeloma cells expressing c-kit in vitro and its mechanism. Methods KM3 cells were treated with Imatinib at different concentrations, and cell growth index were evaltuated by XTT assay, cell cycle by flow cytometry, apoptosis by Annexin V/ PI and DNA ladder, and change in protein level by Western blot. Results Imatinib inhibited proliferation of KM3 cells at concentrations more than 0.25 μmol/L in a dose-dependent manner, and the 48 h IC50 was 0.33 μmol/L （P 〈 0.01 ）. Imatinib arrested cell in G0/G1 phase, Annexin V/PI staining and DNA ladder indicated that Imatinib had a substantial effect on inducing apoptosis of KM3 cells in a dose-dependent manner and induced pro-caspase-3 and poly ADP-ribose polymerase （PARP） cleaved. Imatinib inhibited expression of c-kit and provoked a decrease of IL-6 induced c-kit phosphorylation in vitro. Conclusion Imatinib inhibits KM3 cells proliferation and induces the cells apoptosis by inhibiting c-kit signalling transduction.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2008年第4期230-233,共4页
Chinese Journal of Hematology
基金
广东省科技攻关基金（2006836001008）
