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

Abstract

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.

Circulating Melanoma-Derived Extracellular Vesicles: Impact on Melanoma Diagnosis, Progression Monitoring, and Treatment Response

“…Interestingly, both of these features are related to melanoma initiation and progression. Similarly, mRNA expression of the major histocompatibility antigen HLA-C expression appears to be downregulated in EVs isolated from melanoma cell lines in comparison to normal melanocytes ( HEMa) , suggested as an indicator of poor immunogenicity [ 79 ].…”

Feasibility of tumor‑derived exosome enrichment in the onco‑hematology leukemic model of chronic myeloid leukemia

“…To quantify the RNA extracted from the vesicles, the Y4 transcript was used as an internal reference gene (47,48), and quantified using a 20X TaqMan-MGB-FAM-probe assay for Y4 (5' Y4 RNA) (49). The conventional ABL1 gene was excluded since it is not considered a reliable internal reference gene for exosomes (47,48,(50)(51)(52).…”

Ultraviolet B irradiation enhances the secretion of exosomes by human primary melanocytes and changes their exosomal miRNA profile

“…Cells communicate either via secreted soluble factors or via EVs [4,5]. Melanocytes can release EVs, and researches have used mRNAs or miRNAs contained in exosomes derived from primary melanocytes as a reference to study the contents of melanoma cell-derived exosomes [34,35]. There are relatively few studies that have focused on the effects of melanocytes on other cells.…”