GSE109356
Cellular and extracellular vesicle miRNA profiles in GFP-PC3 and cavin-1-PC3 cell lines.
This study assessed exosomal and cellular miRNA profiles in GFP-PC3 cells and ectopically expressed cavin-1 PC3 cell lines. To understand the consequences of cavin-1 on extracellular vesicle (EV)-contained miRNAs, a small RNA-seq experiment was performed on cells and secreted EVs for GFP-PC3 and cavin-1-PC3 cells. Small RNA were selected for sequencing using the NEBNext Small RNA library Prep Kit. Libraries were size selected using a Perkin Elmer Labchip XT. Whole cell and EV libraries were multiplexed from three biological replicates and sequenced on an Illumina NextSeq500 mid-output run at a read depth of 1million reads per sample. While the raw data was mapped to the human miRNAome using RNA-STAR and miRBase (v21) as reference, 317 mature miRNAs were identified in EVs at varying abundances depending on cell of origin. The 317 miRNAs identified in both cell and EVs were further processed. After RUVseq normalization, DESeq2 analysis was used to identify differences between cellular miRNA profiles and EV miRNA profiles from the two different cell lines. This analysis ultimately defined the repertoire of selectively and passively exported exosomal miRNAs mediated by non-caveolar caveolin-1 and cavin-1 in a prostate cancer cell line. Aim: To assess exosomal miRNA compositional changes evoked by ectopic expression of cavin-1 in the PC3 advanced prostate cancer cell line. Objectives: 1. Determine differential expression of miRNAs in cells and 2. determine differential abundance of miRNAs in extracellular vesicles secreted from cavin-1 expressing or GFP expressing cells. Overall design: miRNA profiles were sequenced from two different cell lines and corresponding extracellular vesicles, in triplicate, on an Illumina NextSeq500