化疗药物对骨髓间充质干细胞的损伤作用

背景：多种化疗药物在杀灭肿瘤细胞的同时会产生骨髓抑制的不良反应,不仅仅损伤造血细胞,也对骨髓间充质干细胞造成损伤,明确各种化疗药物对骨髓间充质干细胞的作用有利于合理选择、搭配化疗药物,实现高效、低毒的临床疗效。目的：探讨不同化疗药物对骨髓间充质干细胞增殖、分化和支持造血能力的影响。方法：获取正常成人骨髓间充质干细胞（n=8）,用不同剂量化疗药物（环磷酰胺、马利兰、阿糖胞苷、柔红霉素、长春新碱、足叶乙甙、甲氨蝶呤、地塞米松）对骨髓间充质干细胞进行处理。检测化疗药物对骨髓间充质干细胞的增殖、凋亡作用及化疗药物撤除后骨髓间充质干细胞的恢复能力;体外诱导药物处理后骨髓间充质干细胞向脂肪细胞和骨细胞分化,通过油红O和Von Kossa染色鉴定;RT-PCR检测药物处理后骨髓间充质干细胞造血因子的表达;集落形成实验检测药物处理后骨髓间充质干细胞支持造血的功能。结果与结论：两种剂量的长春新碱、柔红霉素、足叶乙甙、阿糖胞苷和地塞米松不同程度诱导骨髓间充质干细胞凋亡,并且抑制骨髓间充质干细胞的增殖,其中长春新碱和地塞米松的抑制作用可以逆转,而柔红霉素、足叶乙甙和阿糖胞苷对间充质干细胞的抑制作用持续存在。化疗药物处理后骨髓间充质干细胞仍具有多向分化能力,可以在体外分化为脂肪细胞和成骨细胞。化疗药物处理后骨髓间充质干细胞同正常细胞一样表达造血相关因子：干细胞因子、单核细胞集落刺激因子、白细胞介素6和粒细胞集落刺激因子。柔红霉素、长春新碱、足叶乙甙和阿糖胞苷处理后骨髓间充质干细胞支持造血能力明显下降。结果显示临床常用的一些化疗药物（长春新碱、柔红霉素、足叶乙甙、阿糖胞苷）可以影响骨髓间充质干细胞的增殖和支持造?

Damage of chemotherapy agents to bone marrow mesenchymal stem cells

BACKGROUND： Many chemotherapy drugs can produce adverse reactions, such as bone marrow suppression, when they kill tumor cells. It cannot only damage hematopoietic ceils, but also damage bone marrow mesenchymal stem cells. To understand the role of various chemotherapy agents on bone marrow mesenchymal stem cells is conductive to rational choice of chemotherapy drugs as well as achievement of high-efficiency, low-toxicity, clinical efficacy. OBJECTIVE： To investigate the changes in proliferation, differentiation and hematopoiesis support ability of bone marrow mesenchymal stem cells after exposure to chemotherapy agents. METHODS： Mesenchymal stem cells were isolated from normal adult bone marrow and cultured in media with different chemotherapy agents （cyclophosphamide, busulfan, cytarabine, daunorubicin, vincristine, etoposide, methotrexate, dexamethasone）. After exposure to chemotherapeutic agents, the changes of cell proliferation,apoptosis and recovery ability were detected. In vitro drug-treated bone marrow mesenchymal stem cells differentiated into adipocytes and osteocytes, identified by oil red O and Von Kossa staining. RT-PCR method was used to detect the expression of hematopoietic cytokines in bone marrow mesenchymal stem cells after drug treatment. Colony formation assay was used to detect the hematopoiesis support ability of bone marrow mesenchymal stem cells after drug treatment. RESULTS AND CONCLUSION： Bone marrow mesenchymal stem cells were resistant to three agents： cyclophosphamide, busulfan and methotrexate, and other agents could induce apoptosis of bone marrow mesenchymal stem cells and reduce the proliferation of bone marrow mesenchymal stem cells. However, bone marrow mesenchymal stem cells recovered after withdrawal of vincristine and dexamethasone. Bone marrow mesenchymal stem cells, after exposure to chemotherapy agents, still had the ability of multi-directional differentiation, which could differentiate into adipocytes and osteobiasts in vitro. Moreover, chemotherapy agen