Landscape profiling of small RNAs in extracellular vesicles secreted by senescent human stromal cells after DNA damage treatment

Blooming advancements in next generation sequencing (NGS) have revolutionized system-based analysis of genome-wide expression, cellular pathways and stress responses. We performed this non-coding RNA-associated milestone study to document comprehensively the small RNA molecules carried by extracellular vesicles (EVs) from human stromal cells, which develop typical cellular senescence and manifest the senescence-associated secretory phenotype (SASP) after genotoxic stress in culture. Transcript profiles of primary normal human prostate stromal cells (PSC27) were generated by deep sequencing, in triplicates, using Illumina HiSeq X10. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays.