Autocrine production of TGF-beta confers resistance to apoptosis after an epithelial-mesenchymal transition process in hepatocytes: Role of EGF receptor ligands

Abstract

Transforming growth factor-beta (TGF-beta) induces apoptosis in fetal rat hepatocytes. However, a subpopulation of these cells survives, concomitant with changes in phenotype, reminiscent of an epithelial-mesenchymal transition (EMT). We have previously suggested that EMT might confer cell resistance to apoptosis (Valdés et al., Mol. Cancer Res., 1: 68-78, 2002). However, the molecular mechanisms responsible for this resistance are not explored yet. In this work, we have isolated and subcultured the population of hepatocytes that suffered the EMT process and are resistant to apoptosis (TGF-beta-treated fetal hepatocytes: TbetaT-FH). We prove that they secrete mitogenic and survival factors, as analyzed by the proliferative and survival capacity of conditioned medium. Inhibition of the epidermal growth factor receptor (EGFR) sensitizes TbetaT-FH to die after serum withdrawal. TbetaT-FH expresses high levels of transforming growth factor-alpha (TGF-alpha) and heparin-binding EGF-like growth factor (HB-EGF) and shows constitutive activation of the EGFR pathway. A blocking anti-TGF-alpha antibody restores the capacity of cells to die. TGF-beta, which is expressed by TbetaT-FH, mediates up-regulation of TGF-alpha and HB-EGF expression in those cells. In summary, results suggest that an autocrine loop of TGF-beta confers resistance to apoptosis after an EMT process in hepatocytes, through the increase in the expression of EGFR ligands.