Cantos-Cortes Ana, Amengual Josep, Espinosa-Sotelo Rut, Bertran Esther, Vaquero Javier, de Galarreta Marina Ruiz, Molina-Sanchez Pedro, Serrano Teresa, Ramos Emilio, Calvo Mariona, Scialpi Rosanna, Dituri Francesco, Giannelli Gianluigi, Lujambio Amaia, Gonzalez-Sanchez Ester, Fabregat Isabel
TGF-Beta and Cancer Group - Oncobell Program, Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet de Llobregat, Barcelona, Spain.
CIBEREHD, National Biomedical Research Institute on Liver and Gastrointestinal Diseases, Instituto de Salud Carlos III, Madrid, Spain.
Liver Int. 2025 Oct;45(10):e70351. doi: 10.1111/liv.70351.
Hepatocellular carcinoma (HCC) has a poor prognosis and limited treatment options. TGF-β is a promising therapeutic target, but its dual role, as both a tumour suppressor and promoter, complicates its clinical application. While its effects on tumour cells are increasingly understood, its impact on the tumour stroma remains unclear. This study developed novel in vitro and in vivo mouse models to investigate the role of TGF-β in tumour-stroma interactions under immunocompetent conditions, aiming to evaluate its therapeutic potential in liver cancer.
Mouse liver tumour cell lines were established by hydrodynamic tail vein injection of two different combinations of transposon vector constructions: MYC-LucOS;CTNNB1;SB13 (AL1099 cells) and MYC-Luc;CTNNB1;SB13 (AL1184 cells) to be used in in vitro and in vivo studies.
These cell lines exhibited contrasting phenotypes and responses to TGF-β. We selected AL1099 cells, which displayed an epithelial phenotype and responded to TGF-β with growth inhibition, apoptosis, and induction of Epithelial-Mesenchymal Transition (EMT), and AL1184 cells, which exhibited a mesenchymal-like phenotype and showed enhanced proliferation in response to TGF-β. Both cell lines demonstrated robust growth in 2D and 3D cultures and were co-cultured with RAW 264.7 macrophages, GRX hepatic stellate cells, or both, to explore tumour-stroma interactions. Importantly, AL1099 and AL1184 cells were able to grow in vivo, forming tumours in syngeneic orthotopic models suitable for pharmacological studies.
Together, these findings highlight the value of these models for advancing therapeutic strategies targeting TGF-β, either alone or in combination with immunotherapy, in liver cancer.
肝细胞癌（HCC）预后较差且治疗选择有限。转化生长因子-β（TGF-β）是一个有前景的治疗靶点，但其作为肿瘤抑制因子和促进因子的双重作用使其临床应用变得复杂。虽然其对肿瘤细胞的作用越来越为人所知，但其对肿瘤基质的影响仍不清楚。本研究建立了新的体外和体内小鼠模型，以研究TGF-β在免疫活性条件下肿瘤-基质相互作用中的作用，旨在评估其在肝癌中的治疗潜力。
通过尾静脉高压注射两种不同组合的转座子载体构建物建立小鼠肝肿瘤细胞系：MYC-LucOS;CTNNB1;SB13（AL1099细胞）和MYC-Luc;CTNNB1;SB13（AL1184细胞），用于体外和体内研究。
这些细胞系表现出不同的表型和对TGF-β的反应。我们选择了表现出上皮表型并对TGF-β产生生长抑制、凋亡和上皮-间质转化（EMT）诱导反应的AL1099细胞，以及表现出间充质样表型并对TGF-β产生增殖增强反应的AL1184细胞。两种细胞系在二维和三维培养中均显示出强劲的生长，并与RAW 264.7巨噬细胞、GRX肝星状细胞或两者共同培养，以探索肿瘤-基质相互作用。重要的是，AL1099和AL1184细胞能够在体内生长，在适合药理学研究的同基因原位模型中形成肿瘤。
总之，这些发现突出了这些模型对于推进针对TGF-β的治疗策略（单独或与免疫疗法联合）在肝癌中的价值。
