The availably of 20-hydroxyeicosatetraenoic acid (20-HETE) is required for the metastatic features of human castration resistant prostate cancer PC3 cells

SACCA P; PANELLO LC; NOWICKI S; COLOMBERO C; COSTAS MA

Mar del Plata

Congreso; LXI REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA (SAIC); 2016

Sociedad Argentina de Investigación Clínica

The 20-hydroxylation of arachidonic acid renders 20-HETE.Previous results from our group have shown that the inhibition of20-HETE synthesis reduces androgen-sensitive prostate cancer(PCa) cells viability. The aim of this study was to evaluate therole of 20-HETE on the in vitro characterization of metastaticprocesses in PCa cells. Two human PCa cell lines were used:LNCaP (androgen-sensitive) and PC-3 (androgen-insensitive);both were incubated with either 20-HETE or HET0016 (a selectiveinhibitor of 20-HETE synthesis). Scratch wound assay (cellmigration), and colony growth in soft agar (anoikis resistance)were performed. Also, E-cadherin and vimentin protein expression(epithelial-mesenchymal transition, EMT) were assessed byWB. Cell localization and structure of actin, tubulin and vimentinwere determined by IF. PC-3 cells migration in control conditionswas 38%, this was reduced to 30%* and 25%*** by HET0016 1and 10uM, respectively; also, incubation with 20-HETE 100nMincreased cell migration to 54%***. The analysis of cytoskeletalproteins distribution revealed that HET0016 disorganized actinfilaments throughout PC-3 cells, while tubulin filaments remainedunchanged. Furthermore, HET0016 reduced by 45%** the ability of PC-3 cells to form colonies in soft agar. Interestingly, theanalysis of proteins involved in EMT showed that HET0016increased the expression of E-cadherin in PC-3, as well as,reduced the expression of vimentin, without modifying its intracellulardistribution. Conversely, 20-HETE decreased theexpression of E-cadherin in PC-3 cells and increased theexpression of vimentin. As for LNCaP cells, the incubationwith HET0016 or 20-HETE did not affect cell migration or theexpression of EMT-related proteins. Our results suggest that20-HETE availability is necessary in the steps involved in metastasisof the highly aggressive prostate cancer cell line PC-3.*p