Regulation of amphiregulin gene expression by àŸ-catenin in human hepatocarcinoma cells: a novel crosstalk between FGF19 and the EGFR system

Hepatocellular carcinoma (HCC) is the most prevalent liver tumor and a deadly disease with limited therapeutic options. Dysregulation of cell signalling pathways is a common denominator in tumorigenesis, including hepatocarcinogenesis. The epidermal growth factor receptor (EGFR) signalling system is commonly activated in HCC, and is currently being evaluated as a therapeutic target in combination therapies. We and others have identified a central role for the EGFR ligand amphiregulin (AR) in the proliferation, survival and drug resistance of HCC cells. <i>AR</i> expression is frequently up-regulated in HCC tissues and cells through mechanisms not completely known. Here we identify the ?-catenin signalling pathway as a novel mechanism leading to transcriptional activation of the <i>AR</i> gene in human HCC cells. Activation of ?-catenin signalling, or expression of the T41A ?-catenin active mutant, led to the induction of <i>AR</i> expression involving three specific ?-catenin-Tcf responsive elements in its proximal promoter. We demonstrate that HCC cells expressing the T41A ?-catenin active mutant show enhanced proliferation that is dependent on AR expression and EGFR signalling. We also demonstrate here a novel cross-talk of the EGFR system with fibroblast growth factor 19 (FGF19). FGF19 is a recently identified driver gene in hepatocarcinogenesis and an activator of ?-catenin signalling in HCC and colon cancer cells. We show that FGF19 induced AR gene expression through the ?-catenin pathway in human HCC cells. Importantly, AR up-regulation and EGFR signalling were necessary for the induction of cyclin D1 and cell proliferation by FGF19. Finally, we demonstrate a positive correlation between FGF19 and AR expression in human HCC tissues, therefore validating in clinical samples our experimental observations. These findings identify the AR/EGFR system as a key mediator of FGF19 responses in HCC cells involving ?-catenin signalling, and suggest that combined targeting of FGF19 and AR/EGFR may enhance therapeutic efficacy.