卡博替尼、克唑替尼和奥西替尼对小鼠髓源抑制性细胞亚型和凋亡的影响

目的探讨卡博替尼、克唑替尼和奥斯替尼(AZD9291)对小鼠骨髓或脾脏源性抑制细胞(MDSCs)亚群比例、凋亡及增殖的影响。方法免疫磁珠法分离BALB/c小鼠(雄性)骨髓G-MDSCs和M-MDSCs,分别加入卡博替尼(0.01、0.1、0.3和0.9μmol/L)、克唑替尼(0.2、2、20和200μg/mL)或AZD9291(0.01、0.1、1和10μmol/L)培养24h,采用流式细胞术(FCM)检测3种靶向药物对MDSCs亚型的影响,CCK-8法检测MDSCs增殖;流式细胞分选仪分选小鼠骨髓Gr-1+CD11b+细胞,检测Gr-1+CD11b+细胞凋亡。结果克唑替尼处理组骨髓粒细胞型MDSCs(G-MDSCs)、单核细胞型MDSCs(M-MDSCs)比例均下降(P<0.05);卡博替尼处理组脾脏G-MDSCs比例下降(P<0.05);卡博替尼、克唑替尼处理骨髓MDSCs后早期凋亡比例增加(P<0.05),AZD9291处理的MDSCs凋亡比例无明显变化。结论卡博替尼和克唑替尼可能通过诱导MDSCs凋亡降低MDSCs比例。

Effects of cabozantinib,crizotinib and AZD9291 on subsets and apoptosis of mouse MDSCs

Objective To investigate the effects of cabozantinib,crizotinib and AZD9291 on the percentage of subsets,apoptosis and proliferation of myeloid-derived suppressor cells(MDSCs)from normal mouse bone marrow and spleen.Methods Bone marrow G-MDSCs and M-MDSCs of BALB/c mouse were isolated by immunomagnetic beads and co-cultured with different dosages of cabozantinib,crizotinib and AZD9291 for 24 h.The effects of three targeted drugs on MDSCs subtypes were detected by flow cytometry(FCM),the proliferation of MDSCs was detected by CCK-8;the apoptosis of Gr-1+CD11b+cells was detected by flow cytometry.Results The percentage of spleen G(granulocytic)-MDSCs was significantly decreased after treatment with cabozantinib;the percentage of bone marrow G-MDSCs and M(monocytic)-MDSCs was decreased after treated with crizotinib;early apoptosis rate of bone marrow MDSCs was increased after treated with cabozantinib and crizotinib.There was no obvious change for cell apoptosis after treated with AZD9291.Conclusions cabozantinib and crizotinib may decrease the percentage of MDSCs by inducing apoptosis of MDSCs.