New Hepatocellular Carcinoma (HCC) Primary Cell Cultures as Models for Exploring Personalized Anti-TGF-β Therapies Based on Tumor Characteristics

Transforming growth factor-beta (TGF-beta) plays a dual role in hepatocellular carcinoma (HCC), acting as a tumor suppressor in early stages by inducing cell cycle arrest and apoptosis, and as a promoter in advanced stages by fostering tumor progression, epithelial-mesenchymal transition (EMT), and metastasis. Understanding TGF-beta's role in HCC progression, particularly its impact on tumor-stroma interactions, is crucial for developing personalized therapies. This study aims to clarify TGF-beta function in HCC using patient-derived cell lines and advanced 2D and 3D culture models. Three new cell lines (HLC21, HLC19 tumoral, and HLC19 metastatic) were isolated from HCC patient biopsies, characterizing their phenotypic markers and responses to TGF-beta and its inhibitor, galunisertib. HLC21 cells displayed a mixed epithelial-mesenchymal phenotype, responding to TGF-beta suppressing growth and undergoing EMT, which were inhibited by galunisertib. Conversely, HLC19 tumoral and metastatic cells exhibited mesenchymal phenotypes and were resistant to both TGF-beta suppression and galunisertib effects. In 3D co-cultures with hepatic fibroblasts, TGF-beta inhibitory effects were diminished for responsive cell lines, while resistant lines maintained their non-responsiveness. These findings highlight TGF-beta's dual role in HCC and its influence on tumor-stroma crosstalk, offering valuable models for exploring personalized anti-TGF-beta therapies based on tumor characteristics.