脂肪间充质干细胞分泌的Exosome促进结肠癌细胞系上皮间质转化

Exosomes from human adipose-derived mesenchymal stem cells promote epithelial mesenchymal transition in colon cancer

目的：研究间充质干细胞（mesenchymal stem cell，MSC）来源的Exosome对结肠癌细胞系HCT8上皮间质转化（epithelial mesenchymal transition，EMT）的影响。方法从人脂肪组织中分离出MSC后，对MSC进行培养并传代，并对MSC的分化能力进行鉴定。在人脂肪来源的MSC中提取Exosome后，用透射电子显微镜观察并拍照，并用蛋白质印迹法检测其抗原表达情况。在HCT8培养体系中加入人脂肪来源的MSC分泌的Exosome ，定量PCR和蛋白质印迹法检测上皮和间质转化相关标志物的表达，细胞侵袭和迁移实验检测人脂肪来源的MSC分泌的Exosome对HCT8迁移和侵袭的影响。结果人脂肪来源的MSC具有多系分化能力，人脂肪来源的Exosome直径为40～100 nm ，表达CD63、HSP70和HSP90，下调HCT8上皮相关标志E-cadherin和ZO-1的表达，上调间质相关标志Fibronectin的表达，促进HCT8的迁移和侵袭。结论人脂肪来源的MSC分泌的Exosome可促进结肠癌细胞系HCT8的EMT。

Objective To study the influence of exosome derived from mesenchymal stem cell （MSC） on the epi-thelial mesenchymal transition （EMT） of colon cancer cell line HCT8. Method MSC were separated from human ad-ipose tissue and cultured, of which the differentiation potential was identified. The MSC-derived exosomes were ob-served using TEM, and the antibody expression was detected by western blot. HCT8 cells were co-cultured with MSC-derived exosome. The expression of epithelial and mesenchymal markers was detected by real time PCR and Western blot. Transwell chambers were used in the in vitro migration and invasion assay. Result Human adipose de-rived MSC secreted 40-100 nm particles, which have the typical characteristics of exosomes as expressing CD63, HSP70 and HSP90. With the treatment of MSC-derived exosome, the expression of epithelial related markers E-cad-herin and ZO-1 was down-regulated while the expression of mesenchymal marker Fibronectin was up-regulated, and the migration and invasion capacity of HCT8 cells were enhanced. Conclusion MSC-derived exosomes promotes mi gration of the colon cancer cell line HCT8.