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目的:探讨过氧化物酶体增殖物激活受体γ(PPARγ)激动剂罗格列酮(rosiglitazone,RGZ)对白血病NB4细胞的诱导凋亡作用及其作用机制。方法:以不同浓度的RGZ(20～80μmol/L)作用于体外培养的NB4细胞0、24、48及72h,应用RT-PCR法检测PPARγ的表达水平,MTT法检测细胞生长抑制率,流式细胞术检测细胞凋亡率。应用Western blot检测60μmol/L的RGZ作用不同时间后凋亡抑制蛋白XIAP的表达水平,同时检测不同浓度药物作用不同时间后Caspase-3活性的变化。结果:40μmol/L以上的RGZ可显著抑制细胞的生长及诱导细胞发生凋亡,呈现出明显的量-效与时-效关系。RGZ在诱导细胞凋亡的同时,PPARγ mRNA的表达水平逐渐升高,而凋亡抑制蛋白XIAP的表达水平显著下降。Western blot结果还显示Caspase-3被活化出现20kD亚单位片段。结论:RGZ可以通过PPARγ信号途径抑制NB4细胞的生长并诱导细胞发生凋亡,下调凋亡抑制蛋白XIAP的表达水平以及激活Casapse-3可能是RGZ诱导NB4细胞发生凋亡的重要作用机制之一。
AIM: To investigate the apoptosis-inducing effect of peroxisome proliferator-activated receptor gamma (PPARγ) agonist rosiglitazone (RGZ) on leukemic NB4 cells and its mechanism. Methods: NB4 cells cultured in vitro at different concentrations of RGZ (20 ~ 80μmol / L) for 0,24,48 and 72h, RT-PCR method was used to detect the expression of PPARγ. MTT assay was used to detect the growth inhibition rate and flow cytometry Cytometry detected apoptosis rate. Western blot was used to detect the expression level of XIAP after 60μmol / L RGZ treatment for different time. At the same time, the changes of Caspase-3 activity in different concentrations of drugs were detected. Results: RGZ at 40μmol / L could significantly inhibit cell growth and induce cell apoptosis, showing a dose-response and time-effect relationship. RGZ induced PPARγ mRNA expression in a dose-dependent manner, while XIAP expression decreased significantly. Western blot results also showed that Caspase-3 was activated 20kD subunit fragment. CONCLUSION: RGZ can inhibit the growth of NB4 cells through PPARγ signaling pathway and induce apoptosis. Down-regulation of XIAP expression and activation of Casapse-3 may be one of the important mechanisms of RGZ-induced NB4 cell apoptosis.