Director of Center of Excellence for Stem Cells and Regenerative Medicine Zewail City of Science and Technology October City, Al Jizah, Egypt
Abstract: Hepatocellular carcinoma (HCC) is a leading cause of cancer death. Mesenchymal stromal cells (MSCs) hold promise for HCC therapy. However, controversial reports emerged on MSCs role in cancer. MSCs recruited to hepatic site of injury may undergo metabolic reprogramming and acquire cancer associated fibroblast (CAF)-like characteristics, promoting HCC progression. We investigated MSCs metabolic reprogramming within HCC microenvironment (ME). We studied MSCs role in promoting HCC progression via cellular metabolites exchange between indirectly cocultured bone marrow MSCs (BMMSCs) and Huh7 HCC cell line (Huh7). We evaluated cellular bioenergetics with a focus on lactate monocarboxylate transporters (MCT1&4). Genotypic analysis of cocultured BMMSCs showed upregulation in CCL2, cMYC, FGFR1, IL8, αSMA, MMP2, SDF1, Vimentin, VEGFα CAF markers, MCT4 and MCT1 downregulation. Concomitant with increase in cellular proliferation as assessed by MTT assay, cell cycle analysis by flowcytometry and genotypic analysis by qRTPCR. AnnexinV/propidium iodide staining in cocultured Huh7 showed downregulation in apoptosis markers, Bcl2 antiapoptotic gene and upregulation of HTERT, Bax and Caspase3 apoptotic genes. Increase in cell migration was observed in cocultured Huh7 using scratch assay, upregulation of epithelial mesenchymal transition genes, Snail, Slug, Vimentin and downregulation of Ncadherin. Genotypic analysis of cocultured Huh showed downregulation of ALDOC, Glut1, ERRᵞ and PKM2, upregulation of ENO1, TP1, PGK1, IDH1 glycolytic genes and upregulation of ATP6v, Cox11, NDUF5&10 oxidative phosphorylation genes. Along with MCT4 downregulation and MCT1 upregulation, which was confirmed by western blot. Biochemical analysis of glucose consumption, lactate, pyruvate, hydrogen peroxide, lactate dehydrogenase, reactive oxygen species and urea showed downregulation in cocultured Huh7 and upregulation in cocultured BMMSCs. Our data show that HCCME may induce BMMSCs metabolic reprograming into CAF-like cells promoting HCC progression, which is mediated via dysregulation of intracellular gene expression of metabolites and their associated metabolic pathways, mainly achieved via lactate transporters. MSCs fate in HCCME thus needs further evaluation and their putative HCC therapeutic use.
Funding Source: Grant #46721 from the Egyptian Science and Technology Development Fund, Cairo, Egypt Grant#5275 from Academy of Scientific Research and Technology, Cairo, Egypt
