白血病成骨细胞屏蔽共培养AML细胞化疗杀伤作用的研究

目的探讨白血病成骨细胞对急性髓性白血病(AML)细胞屏蔽化疗杀伤的作用,及其与促血小板生成素/血小板生成素受体(TPO/c-MPL)信号通路的关系。方法分离培养白血病骨髓间充质干细胞并行成骨诱导,ELISA法检测成骨细胞分泌血小板生成素(TPO)水平,流式细胞仪检测人红白血病细胞株(HEL)c-MPL表达。成骨细胞与HEL细胞共培养,观察在化疗药物柔红霉素干预下HEL细胞存活率。结果白血病骨髓成骨细胞培养d7、d14 TPO表达分别为(176.84±15.32)ng/mL和(198.24±13.49)ng/mL,明显高于正常对照的TPO水平,分别为d7(140.13±20.12)和d14(157.66±18.82)ng/mL。应用柔红霉素干预后,AML成骨细胞与HEL细胞共培养组半抑制浓度(IC50)为(3.320±0.218)μg/mL,高于HEL+正常来源成骨细胞组(2.236±0.167)μg/mL和HEL+hFOB1.19组(2.254±0.111)μg/mL。生长曲线研究发现,在不同浓度DNR干预下,AML成骨细胞与HEL细胞共培养组中HEL细胞的细胞存活率最大,均高于其他HEL共培养组。结论白血病成骨细胞能有效屏蔽化疗药物对白血病细胞的杀伤作用,这种作用可能与白血病骨髓微环境内TPO/c-MPL信号通路相关。

Function and mechanism study of the leukemia osteoblast cells preventing the co-cultured AML cells from chemotherapy

Objective To study the role of leukemia osteoblasts on shielding the chemotherapy killing of acute myeloid leukemia cells and the relationship between leukemia osteoblasts and TPO/c-MPL signaling pathway.Methods Isolated and co-cultured leukemia bone marrow mesenchymal stem cells were induced into osteoblasts in vitro,the secretion of thrombopoietin of osteoblasts was detected by ELISA and the expression of c-PL of HEL cells was detected by flow cytometry.The osteoblasts and HEL cells were co-cultured in vitro,and the survival rate of HEL cells that intervented by daunorubicin were observed.Results The TPO expression of leukemia bone marrow osteoblasts was significantly higher than the control group,the osteoblasts that induced by leukemia cells shielded the HEL cells in chemotherapy killing was significantly stronger than that of the control group after intervented by daunorubicin.Conclusion Leukemia osteoblasts can shield the chemotherapy killing of acute myeloid leukemia cells,and this may have relationship with TPO/c-MPL signaling pathway.