小鼠黑素瘤细胞系分泌的外泌体促进间充质基质细胞增殖和迁移

摘要

Objective To investigate the interactions between melanoma-derived exosomes and the microenvironment.Methods The exosomes were isolated from the culture medium of mouse melanoma cells and then co-cuhured with mesenchymal stromal cells (MSC) after identification.Immunofluorescence assay was performed to observe the exosomes engulfed by MSC.CCK-8 and transwell assays were used to evaluate the proliferation and migration of MSC.Effects of the exosomes on the expression of α-smooth muscle actin (α-SMA) in MSC were analyzed by Western blot.Results Co-culture of MSC with melanoma cell-derived exosomes enhanced the proliferation and migration of MSC as well as the expression of α-SMA.All of the changes mediated by the exosomes could be blocked by using the inhibitor of TGF-β receptor.Conclusion Melanoma cell-derived exosomes enhanced the proliferation and migration of MSC as well as the expression of α-SMA through TGF-β signaling pathway,which provided an advantageous microenvironment for melanoma progression.%目的 研究黑素瘤来源的外泌体(exosome)如何与微环境相互作用.方法 分离并鉴定小鼠黑素瘤细胞培养液上清中的外泌体,并与间充质基质细胞(MSC)共培养.利用免疫荧光观察细胞摄入外泌体情况,CCK-8及transwell实验测定MSC增殖与迁移的变化情况,Western blot检测外泌体对MSC的α平滑肌肌动蛋白(α-SMA)表达的影响.结果 发现MSC在摄取外泌体后,增殖和迁移能力显著上升,α-SMA的表达明显上调.且这些变化都可被TGF-β受体的抑制剂阻断.结论 外泌体可能通过TGF-β诱导MSC构建微环境,促进黑素瘤的生长与转移.