Abstract：
Exosomes are small membrane-derived vesicles that transmit DNA constituents, mRNAs, microRNAs, and proteins from donor cells to a receiver cell. Various cells comprising of mesenchymal, immune, and cancer cells discharge exosomes. Cancer cell exosomes form the entry and reprogramming of essentials connected to a tumor environment. Melanoma-derived exosomes transport diverse proteins such as c-MET and RAB27a, which leave a melanoma mark. Increased mesenchymal epithelial transition (MET) expressions in serum exosomes have been considered an indicator of disease progression. Meanwhile, RAB27a has been identified as being involved in exosome discharge and trafficking. Decreased expressions of RAB27a in human melanoma cells have shown to diminish exosome release.We examined the effects of the downregulation and upregulation of RAB27a and c-MET in human dermal fibroblasts by utilizing the isolated exosomes of malignant melanoma cell lines. Melanoma exosomes derived from cancer cells conveyed information to healthy dermal fibroblasts and stem cells while inducing phenotypic change. In this chapter, we show optimized protocols that were used by our group for in vitro analysis with melanoma exosomes.
摘要：
外泌体是传递DNA成分的小膜来源的囊泡，mRNA,microRNAs,和蛋白质从供体细胞到接受细胞。包含间充质的各种细胞，免疫，癌细胞排出外泌体。癌细胞外泌体形成与肿瘤环境相关的要素的进入和重编程。黑色素瘤来源的外泌体转运多种蛋白质，如c-MET和RAB27a，留下黑色素瘤痕迹.血清外泌体中的间充质上皮转化（MET）表达增加被认为是疾病进展的指标。同时,RAB27a已被确定参与外泌体排放和贩运。RAB27a在人黑素瘤细胞中的表达降低已显示减少外泌体释放。我们通过利用分离的恶性黑色素瘤细胞系的外泌体，研究了人真皮成纤维细胞中RAB27a和c-MET的下调和上调的作用。来自癌细胞的黑色素瘤外泌体向健康的真皮成纤维细胞和干细胞传递信息，同时诱导表型变化。在这一章中,我们展示了我们小组用于黑色素瘤外泌体体外分析的优化方案.
