20-Hydroxyeicosatetraenoic Acid (20-HETE), a Cytochrome P450 (CYP) Metabolite of Arachidonic Acid, Contributes to the Metastatic Features of Human Castration-Resistant Prostate Cancer Cells

NOWICKI SUSANA; PANELO LAURA C; COSTAS MÓNICA A,; COLOMBERO CECILIA; SACCA PAULA

Orlando

Congreso; 99th Annual Meeting & Expo of The Endocrine Society; 2017

Endocrine Society

The hydroxylation of arachidonic acid by 20-hydroxylases of the cytochrome P450 family (CYP4F2 and CYP4A11) renders 20-hydroxyeicosatetraenoic acid (20-HETE), which has been implicated in oncogenic characteristics in several types of tumors. Previous results from our group have evidenced the expression of both, CYP4F2 and CYP4A11, in two human prostate cancer cell lines: LNCaP (androgen-sensitive) and PC-3 (androgen-insensitive) and have shown that the inhibition of 20-HETE synthesis reduces the viability only in androgen-sensitive prostate cancer cells.The aim of this study was to evaluate the role of 20-HETE on the metastatic features in vitro in prostate cancer cells.LNCaP and PC-3 cells were incubated with either 20-HETE or HET0016 (a selective inhibitor of 20-HETE synthesis). Scratch wound assay (cell migration), and colony growth in soft agar assay (anoikis resistance) were performed. Also, E-cadherin and vimentin protein expression (epithelial-mesenchymal transition, EMT) were assessed by Western Blot. Additionally, the release of matrix metalloproteinase-2 (MMP-2), involved in the degradation of extracellular matrix, was assessed by zymography assay. Intracellular localization and structure of actin, tubulin and vimentin were determined by immunofluorescence. Results were analyzed using one-way ANOVA followed by Dunnet?s post-hoc analysis.The analysis of proteins involved in EMT showed that HET0016 10uM increased by 80% (p