c-myc抑制剂10058-F4对Ph^+ALL细胞凋亡的影响及相关机制研究

Effect of c-myc inhibitor 10058-F4 on the apoptosis of Ph^+ acute lymphoblastic leukemia cells and the underlying mechanisms

目的探讨c-myc抑制剂10058-F4对Ph^+急性淋巴细胞白血病(Ph^+ALL)细胞Sup-B15的作用及其分子调控机制,为将10058-F4开发成为治疗Ph^+ALL药物提供理论依据。方法流式细胞术检测10058-F4对Sup-B15细胞凋亡和细胞周期的影响,以及对Ph^+和Ph^-的原代ALL细胞凋亡和坏死的影响;Western blotting检测10058-F4对c-myc及其下游调控基因蛋白表达的影响。结果 10058-F4显著地诱导Sup-B15细胞凋亡,但对细胞周期无明显影响;10058-F4分别诱导Ph^+和Ph^-的原代ALL细胞发生明显的凋亡和坏死。40μmol/L的10058-F4显著抑制c-myc、Bcl-xl、Mcl-1和Bcl-abl的蛋白表达,但对Bcl-2、Bax和p-crkl蛋白表达均无明显影响。阳性对照药物PPP和IM均显著抑制c-myc、Mcl-1和p-crkl蛋白的表达,同时,PPP也明显地抑制Bcl-2蛋白的表达。结论 10058-F4能有效促进Ph^+的Sup-B15细胞系和原代细胞的凋亡,其机制与抑制c-myc及其调控的抗凋亡蛋白Bcl-2和Bcl-xl的表达相关。

Objective To investigate the effect of c-myc inhibitor 10058-F4 on philadelphia chromosome positive (Ph^+) acute lymphoblastic leukemia (Ph^+ALL) cells and its underlying mechanisms.Methods Flow cytometry was performed to detect the apoptosis and cell cycle distribution of Ph^+ALL cell line Sup-B15 cells,and the apoptosis and necrosis of primary cells from Ph^+ and Ph - ALL patients after 10058-F4 treatment,respectively.Western blotting was used to detect the effect of 10058-F4 on the protein expression of c-myc and its downstream regulatory molecules.Results 10058-F4 induced significant apoptosis of of Sup-B15 cells.but had no effect on cell cycle,while 10058-F4 also significantly induced the apoptosis and necrosis in primary Ph^+ and Ph^- ALL cells.40 μmol/L 10058-F4 significantly inhibited the protein expression of c-myc,Bcl-2,Bcl-xl,Mcl-1 and Bcl-abl,but had no effect on the protein expression of Bcl-2,Bax and p- crkl.As the positive control drugs,PPP and IM markedly reduced the protein expression of c-myc,Mcl-1 and p- crkl,and PPP also apparently repressed the expression of Bcl-2 protein.Conclusion 10058-F4 can effectively promote the apoptosis of Ph^+ Sup-B15 cells and primary ALL cells,which is related to inhibition of the expression of c-myc and its downstream regulatory anti-apoptotic proteins Bcl-2 and Bcl-xl.