Spermidine synthase regulates extracellular vesicle secretion in castration resistant prostate cancer.

Extracellular vesciles (EVs) have an important role in the tumor progression. However, the precise regulatrory mechanisms of EV secretion has not been clarified yet. We performed quantitive high-throughput screening to eluciade the key miRNAs for EV biogenesis in castration resistant prostate cancer (CRPC) using 22RV1 cells, and miR-1908 was selected as the miRNA regulating EV biogenesis in CRPC. To identify the genes that could be targeted by miR-1908, we performed mRNA micorarray analysis in 22RV1 after transfected of miR-1908 mimic or control miRNA. 22RV1 cells were seeded into 6-well plates, and 48 hours later, the transfection of miR-1908 mimic or controls was accomplished with the DharmaFECT Transfection Reagent 1 (Dharmacon) according to the manufacturer’s protocol. After 24 hours, the CM was changed to Advanced DMEM medium containing 2 mM L-glutamine without an antibiotic-antimycotic. Total mRNAs were extracted from the 22RV1 cells 48 hours after the medium change. We used Agilent microarray chip (SurePrint G3 Human GE v3).