Glycosylated 4-methylumbelliferone reduces the expressions of cancer stem cells in hepatocellular carcinoma

CAECILIA SUKOWATI; MARIA MOLEJON; ALINA GONZALEZ; CLAUDIO TIRIBELLI; GISELA WEIZ

Congreso; EASL Liver Cancer Summit 2023; 2023

Background and Aims: The inhibition of hyaluronic acid (HA) may block the growth of various cancers, including hepatocellular carcinoma (HCC). The 4-methylumbelliferone (4-MU), an HA enzymatic inhibitor, was shown to reduce hepatic fibrosis, inflammation, and the presence of cancer stem cells (CSC). Recently, we demonstrated that a glycosylated 4-MU, namely 4-MUR, with the addition of rhamnose residue in 4-MU, constituted a better strategy to target HCC cells while sparing normal liver cells due to the interaction with the rhamnose-specific asialoglycoprotein receptor in HCC cells. This study aimed to investigate the comparison between 4MUR and 4MU to the profile of the CSC profiles of HCC.Method: The profile of CSC markers in HCC cell lines JHH6 and Huh7 upon treatment of 4-MU and 4- MUR was evaluated using flow cytometer and real-time PCR for counting of the cells and gene expression, respectively. The cytotoxicity of the compounds was assessed by MTT assay. Immortalized hepatocytes cell line IHH was used as control.Results: 4-MUR was toxic only in HCC cell lines and not in immortalized hepatocytes. 4-MUR treatment better reduced the number of cells expressing CSC markers EpCAM and CD13/ANPEP compared to 4- MU in both HCC cell lines. Reduction of CD133/Prom-1 was also noticed in Huh7 which expresses this marker. Flow cytometric data was then compared with that in gene expression, showing concordant results. Interestingly, the 4-MUR did not reduce the expressions of both CD44, a receptor of HA, and CD90/THY-1 mRNA in IHH cells.Conclusion: 4-MUR capacity to reduce the expressions of CSCs in HCC cells was comparable, if not better, to those of 4-MU. Importantly, 4-MUR was selective to target only HCC cells while being less efficient in normal hepatocytes.