The aging effect of chemotherapy on cultured human mesenchymal stem cells
The aging effect of chemotherapy on cultured human mesenchymal stem cells
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DOI：
10.1016/j.exphem.2011.08.009
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发表时间：
2011-12-01
影响因子：
2.6
通讯作者：
Tarella, Corrado
中科院分区：
文献类型：
--
作者：
Buttiglieri, Stefano;Ruella, Marco;Tarella, Corrado
包括化学治疗药物在内的各种试剂可以诱导细胞衰老。然而，衰老途径中涉及的机制，特别是化疗对端粒的压力，仍然不确定。为了解决这些问题，将人间充质干细胞（MSC）作为靶细胞进行评估，以研究化疗引发的衰老过程。从正常成人的骨髓（BM）细胞获得MSC，并在血小板裂解物的存在下生长。对培养的MSC进行免疫表型、生长和分化特性鉴定。将MSC暴露于10 nM阿霉素和500 ng/mL依托泊苷，亚致死剂量诱导DNA双链断裂。端粒长度（TL）进行了评估，通过流式荧光原位杂交和Southern印迹。在药物暴露后5天，在MSC中可检测到初始TL缩短，与培养中的未处理细胞相比，在7、14、21和28天进行性减少。单次暴露后，MSC在培养中长达28天无法恢复丢失的端粒序列。在药物暴露后早期记录了ATM磷酸化，而没有观察到端粒酶激活。化疗诱导的TL缩短与体外克隆形成活性降低和脂肪分化加速有关。在自然老化的MSC中观察到分化模式的类似行为。这些结果表明，培养的MSC代表了一个有用的细胞模型，研究新的药物，可能有利于或相反，可能会防止TL损失的人干细胞。TL缩短是先前化疗介导的DNA损伤的永久标志，并预测受损的增殖和分化潜力。(C)2011 ISEH -血液学和干细胞学会。爱思唯尔公司出版
Various agents, including chemotherapeutic drugs, can induce cell senescence. However, the mechanisms involved in the aging pathway, particularly the stress that chemotherapy imposes on telomeres, are still undefined. To address these issues, human mesenchymal stem cells (MSCs) were assessed as target cells to investigate the initiation of the aging process by chemotherapy. The MSCs were obtained from bone marrow (BM) cells from normal adults and grown in the presence of platelet lysates. Cultured MSCs were identified for immunophenotype, and for growth and differentiation properties. The MSCs were exposed to 10 nM doxorubicin and 500 ng/mL etoposide, sublethal doses that induce DNA double-stranded breaks. Telomere length (TL) was assessed by flow-fluorescence in situ hybridization and Southern blotting. Initial TL shortening was detectable in MSCs at 5 days after drug exposure, with progressive reduction compared with untreated cells at 7, 14, 21, and 28 days in culture. After a single exposure, MSCs were unable to regain the lost telomere sequences for up to 28 days in culture. The ATM phosphorylation was documented early after drug exposure, while no telomerase activation was observed. Chemotherapy-induced TL shortening was associated with reduced clonogenic activity in vitro and accelerated adipose differentiation. Analogous behavior in the differentiation pattern was observed in naturally aged MSCs. These results indicate that cultured MSCs represent a useful cellular model to investigate novel drugs that may favor or, conversely, might prevent TL loss in human stem cells. The TL shortening is a permanent signature of previous chemotherapy-mediated DNA damage, and predicts impaired proliferative and differentiation potential. (C) 2011 ISEH - Society for Hematology and Stem Cells. Published by Elsevier Inc.