miR-26a regulate the extracellular vesicle secretion from prostate cancer cells via targeting PFDN4, CHORDC1 and SHC4. miR-26a regulate the extracellular vesicle secretion from prostate cancer cells via targeting PFDN4, CHORDC1 and SHC4

Extracellular vesicles (EVs) are known to be involved in inter-cellular communication during cancer progression, however, the biogenesis of EVs in cancer cells is not completely unveiled. It has been shown that microRNAs (miRNAs) regulated variety of physiological and pathological phenomena, thus, miRNAs could regulate the EV secretion. Here, we have performed high throughput miRNA-based screening to identify the regulators of EV secretion using ExoScreen assay. By this miRNA-based screening, we identified miR-26a, which was reported as tumor suppressive miRNA, was found to be an miRNA involved in EV secretion from prostate cancer (PCa) cells. To study the effect of miR-26a on gene expression in EV secretion, transcriptome analysis for miR-26a overexpressing PCa cell lines was performed. Overall design: Transfections of 10 nM miR-26a mimics or negative control miRNA (miRNA mimic Negative Control #1, Ambion) were accomplished with the DharmaFECT Transfection Reagent 1 according to the manufacturer’s protocol. After 24 hours, the conditioned medium was changed to Advanced RPMI medium containing an antibiotic-antimyotic and 2mM L-glutamine. Forty-eight hours after the medium change, total RNAs were extracted from them. We used Agilent microarray chip (SurePrint G3 Human GE v3).