血小板活化白血病细胞L1210中AKT及ERK信号通路并降低其对多种药物的敏感性

Platelets Decrease the Sensitivity of Leukemia Cell L1210 to Multiple Drugs via Activiting The AKT and ERK Signalling Pathway

目的:探索血小板对白血病细胞胞内信号通路及药物敏感性的可能影响。方法:分离小鼠血小板,应用流式细胞术观察血小板与白血病细胞L1210共孵育后是否发生相互作用;使用Western blot印迹法检测血小板对白血病细胞L1210胞内信号通路的影响;在共培养体系中分别加入甲氨蝶呤、长春新碱、多柔比星,通过细胞增殖活性试验检测血小板对白血病细胞L1210药物敏感性的影响。结果:新鲜分离的以及固定后的血小板都能与白血病细胞相互结合,并都能上调白血病细胞内AKT以及ERK的磷酸化水平。此外,血小板还显著降低白血病细胞L1210对三种化疗药物的敏感性。结论:血小板可与白血病细胞相互作用,激活白血病细胞AKT和ERK信号通路,并可能通过该途径降低白血病细胞L1210对多种药物的敏感性。

Objective：To explore the effect of platelets on signal transducers and the sensitivity of leukemia cells to chemotherapeuticl drugs in leukemia cells L1210.Methods：Murine platelets were prepared and cocultured with leukemia L1210 cells,and the aggregation between them was observed by flow cytometry.The levels of several transducer proteins in leukemia cells were analyzed with Western blot.In some experiments,methotrexate,vincristine or doxorubicin was added to the coculture system and the cell proliferation was measured by using CCK8 colorigenic methods to detect the sensitivity of leukemia cells to the therapeuticals drugs.Results：Platelets,either freshly prepared or fixed with1%paraformaldehyde,aggregated with leukemia cells.Both fresh and fixed platelets increased the level of phosphorylation of AKT and ERK in leukemia cells as measured with Western blot.Also,platelets significantly decreased the sensitivity of 3 therapeutics to L1210 cells.Conclusion：Platelets may bind with L1210 cells and decrease the sensitivity of the leukemia cells to chemotherapeutics,possibly by activating the AKT and ERK signaling pathways.