p63 induces tumor progression in bladder cancer

ZAMBRANO, MACARENA; ; SCIACCA, MARINELA; ; BELGOROSKY, DENISE; ; LANGLE, YANINA; ; BALARINO, NATALIA; ; SANDES, EDUARDO; ; EIJAN ANA MARIA; LODILLINSKY, CATALINA

CIUDAD AUTONOMA DE BUENOS AIRES

Congreso; IV International Congress in Traslational Medicine. 27-28 de Marzo de 2018, FFyB-FMed-UBA; 2018

Based in the need to develop more precise therapies, bladder cancer has been classified related to its gene expression pattern, in three main groups, similar to those in breast cancer: luminal like, basal like and p53 like bladder tumors(Choi, 2014; Dambauer, 2014). The biological and clinical significance of these signatures remains unclear.p63 characterize basal like bladder tumors, however the p63-related molecular pathways in this pathology are not well known yet.Our hypothesis is that p63 acts as a protumoral factor, by activation of invasive signaling programs. Knocking down for p63 in human bladder cancer cell lines MGHU3 and UMUC14 was developed using a TET-ON system where the p63 depletion is induced externally under doxycycline (DOX). Cells were treated for 96 hours with DOX and p63 expression was analyzed by western blot. We could observe an almost complete abrogation of p63 expression under DOX in MGHU3 and a 60% of depletion in UMUC14 cells. Gelatin degradation assay was performed in both cell lines. We could observe that while DOX has no effect in the degradation ability of Non-Targeting cells, shp63_i cells under DOX degraded 60 percent less compared without DOX. (p≤0,01; ANOVA Tukey between with and without DOX). Migratory ability was tested using the wound healing assay in UMUC14 cells. p63 depleted cells presented a significantly lower potential of 70% to close the wound 24 h after initiation of the wound healing assay (p < 0.0001; t test Mann Whitney, DOXY vs untreated shp63_i cells).Spheroids of both cell lines were grown during 30 days and diameter was compared between with and without DOX (ANOVA with Tukey comparisons ***p