RNA-Seq analysis using WA-treated 22Rv1 human prostate cancer cells revealed upregulation and downregulation of 6993 and 6607 genes, respectively when compared to the vehicle-treated control. The pathways with significantly upregulated gene expression following WA treatment included mitogen-activated protein kinases, focal adhesion, endocytosis, and autophagy as revealed by KEGG and gene ontology analyses. WA treatment also resulted in downregulation of genes associated with metabolic pathways, oxidative phosphorylation, tricarboxylic acid cycle, cell cycle, and base excision repair. This study identifies novel mechanistic targets of WA in prostate cancer. Overall design: Examination of altered genes expression in 22Rv1 human prostate cancer cell line after 16-h treatment with vehicle-treated control (n=3) or 2 µM of WA (n=3)