Liver-derived mesenchymal stem cells promote epithelial-mesenchymal transition in a hepatocellular carcinoma spheroid coculture model

Mesenchymal stem cells (MSC) play a highly debated role in cancer. Potent anti- and pro-tumour properties have been shown in vitro and in vivo depending on the tumour studied and the MSC used. Here, we investigate the interaction between liver-derived MSC and hepatocellular carcinoma (HCC). We postulate that this shared origin can lead to unique insights on the potential implication of MSC in carcinogenesis. We established a 3D coculture spheroid model by combining Human Adult Liver Progenitor Cells (HALPC), liver-derived mesenchymal stem cells described by our team, and one of three human hepatocellular carcinoma cell line (HuH7, HepG2, Hep3B). Cancer cell proliferation was studied through spheroid growth follow-up, chemiluminescent measurement of ATP content and Ki67 immunostaining in light sheet fluorescent microscopy. Cancer cell invasiveness was investigated using transwell and spheroid invasion assays. Total RNA sequencing was conducted on HALPC and cancer cells extracted from control and coculture spheroids. HCC coculture with HALPC resulted in an inhibition of cancer cell proliferation (up to 66% reduction in Ki67-positive cells in coculture ; p = 4.6x10-5), while invasion studies demonstrated the induction of an invasive phenotype. Comprehensive transcriptomic analysis showed massive changes in expression levels in both HALPC and cancer cells following coculture. In cancer cells, gene set enrichment analysis revealed dysregulation of cancer hallmarks gene sets, with an important positive enrichment of the epithelial-mesenchymal transition (EMT) gene set (Normalized Enrichment Score: + 2.7 ; p adj. 1.1x10-18). We showed in a 3D coculture model that liver-derived MSC decreased the proliferation and induced the invasiveness of several human HCC cell lines. This was associated with the induction of an EMT phenotype at the transcriptomic level, shedding additional light on the role of organ resident MSC in carcinogenesis. Ongoing work will confirm the EMT phenotype at the protein level and validate results in vivo.

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de Bodt, G., Ravau, J., Evraerts, J., Stephenne, X., Najimi, M., & Sokal, E. (2025). Liver-derived mesenchymal stem cells promote epithelial-mesenchymal transition in a hepatocellular carcinoma spheroid coculture model. 57th Annual Meeting of the European Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN), Helsinki, Filand. https://hdl.handle.net/2078.5/242114