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Detection of Inflammation-Related Melanoma Small Extracellular Vesicle (sEV) mRNA Content Using Primary Melanocyte sEVs as a Reference

机译:使用原发性黑素细胞sEVs作为参考检测与炎症相关的黑素瘤小细胞外囊泡(sEV)mRNA含量

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摘要

Melanoma-derived small extracellular vesicles (sEVs) participate in tumor pathogenesis. Tumor pathogenesis is highly dependent on inflammatory processes. Given the potential for melanoma sEVs to carry tumor biomarkers, we explored the hypothesis that they may contain inflammation-related mRNA content. Biophysical characterization showed that human primary melanocyte-derived sEVs trended toward being smaller and having less negative (more neutral) zeta potential than human melanoma sEVs (A-375, SKMEL-28, and C-32). Using primary melanocyte sEVs as the control population, RT-qPCR array results demonstrated similarities and differences in gene expression between melanoma sEV types. Upregulation of pro-angiogenic chemokine ligand CXCL1, CXCL2, and CXCL8 mRNAs in A-375 and SKMEL-28 melanoma sEVs was the most consistent finding. This paralleled increased production of CXCL1, CXCL2, and CXCL8 proteins by A-375 and SKMEL-28 sEV source cells. Overall, the use of primary melanocyte sEVs as a control sEV reference population facilitated the detection of inflammation-related melanoma sEV mRNA content.
机译:黑色素瘤衍生的小细胞外囊泡(sEVs)参与肿瘤的发病机理。肿瘤发病机理高度依赖于炎症过程。考虑到黑素瘤sEV携带肿瘤生物标志物的潜力,我们探讨了它们可能包含炎症相关mRNA含量的假设。生物物理特征表明,与人黑素瘤sEV(A-375,SKMEL-28和C-32)相比,人原发性黑素细胞衍生的sEV趋向于更小且具有负(更中性)的ζ电位。使用原代黑色素细胞sEV作为对照人群,RT-qPCR阵列结果证明了黑色素瘤sEV类型之间基因表达的相似性和差异。最一致的发现是A-375和SKMEL-28黑色素瘤sEV中促血管生成趋化因子配体CXCL1,CXCL2和CXCL8 mRNA的上调。这与A-375和SKMEL-28 sEV源细胞对CXCL1,CXCL2和CXCL8蛋白的生产增加平行。总体而言,使用原代黑素细胞sEV作为对照sEV参考人群有助于检测炎症相关的黑色素瘤sEV mRNA含量。

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