NEW INSIGHTS INTO THE MOLECULAR MECHANISMS THAT PROVOKE THE DEVELOPMENT OF PRENEOPLASTIC ALTERATIONS IN A HEPATOCYTE CELL LINE

CAMPISANO SE; BERTRAN E; LA COLLA A; ECHARTE SM; FABREGAT I; CHISARI AN

Buenos Aires

Congreso; Reunión conjunta de Sociedades de Biociencias; 2017

Previously, it was demonstrated that the lack of aminoacids (Aa)in the culture medium of an immortalized mice hepatocyte cell line(parental line = P) induces apoptosis, and that cells that are ableto survive (selected line = S) showed a more proliferative phenotypeand resistance to apoptotic stimuli, reminiscent of a preneoplasticphenotype. Recent data proposes an essential role for theNADPH oxidase NOX4 as a liver tumor suppressor, so the aim ofthis work was to deep into the molecular mechanisms that supportsuch preneoplastic alterations focusing on growth factors, inflammatorysignals and NOX4, in the cross-roads of these pathways.Cell proliferation was measured using violet crystal, at 0, 24 and48h of cell culture. Different concentrations of cells were employed.We performed qPCR to quantify the expression of NOX4, TGF-β,TGF-α, CXCL-1 and CXCL-2. The levels of NOX4 and pEGFR proteinswere determined by Western Blot. S showed higher proliferationvs P, which was more marked at 48h and using the highest concentrationof cells per well (10 x106) (P vs S, mean±SD: 718±42.77vs 1317±130.8; p