5-azacytidine downregulates the proliferation and migration of hepatocellular carcinoma cells in vitro and in vivo by targeting miR-139-5p/ROCK2 pathway

Tonon, F. et al. (2022) 5-azacytidine downregulates the proliferation and migration of hepatocellular carcinoma cells in vitro and in vivo by targeting miR-139-5p/ROCK2 pathway. Cancers, 14(7), 1630. (doi: 10.3390/cancers14071630) (PMID:35406401) (PMCID:PMC8996928)

Abstract

Background: For hepatocellular carcinoma (HCC), effective therapeutic approaches are lacking. As aberrant gene methylation is a major contributor to HCC development, demethylating drugs such as 5-azacytidine (5-Aza) have been proposed. As most 5-Aza mechanisms of action are unknown, we investigated its phenotypic/molecular effects. Methods: 5-Aza effects were examined in the human HCC cell lines JHH-6/HuH-7 and in the rat cell-line N1-S1. We also employed a xenograft mouse model (HuH-7), a zebrafish model (JHH-6), and an orthotopic syngeneic rat model (N1-S1) of HCC. Results: 5-Aza downregulated cell viability/growth/migration/adhesion by upregulating miR-139-5p, which in turn downregulated ROCK2/cyclin D1/E2F1 and increased p27kip1, resulting in G1/G0 cell accumulation. Moreover, a decrease in cyclin B1 and an increase in p27kip1 led to G2/M accumulation. Finally, we observed a decrease in MMP-2 levels, a stimulator of HCC cell migration. Aza effects were confirmed in the mouse model; in the zebrafish model, we also demonstrated the downregulation of tumor neo-angiogenesis, and in the orthotopic rat model, we observed impaired N1-S1 grafting in a healthy liver. Conclusion: We demonstrate for the first time that 5-Aza can impair HCC development via upregulation of miR-139-5p, which in turn impairs the ROCK2/cyclin D1/E2F1/cyclin B1 pro-proliferative pathway and the ROCK2/MMP-2 pro-migratory pathway. Thus, we provide novel information about 5-Aza mechanisms of action and deepen the knowledge about the crosstalk among ROCK2/cyclin D1/E2F1/cyclin B1/p27kip1/MMP-2 in HCC.

• Item Type: Articles
• Keywords: MMP-2, 5-azacytidine, E2F1, ROCK2, migration, miR-139-5p, hepatocellular carcinoma, cell cycle.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Olson, Professor Michael

Creator Roles

Olson, M.: Conceptualization, Resources, Writing – review and editing

• Authors: Tonon, F., Cemazar, M., Kamensek, U., Zennaro, C., Pozzato, G., Caserta, S., Ascione, F., Grassi, M., Guido, S., Ferrari, C., Cansolino, L., Trotta, F., Kuzmanov, B. G., Forte, G., Martino, F., Perrone, F., Bomben, R., Gattei, V., Elvassore, N., Murano, E., Truong, N. H., Olson, M., Farra, R., Grassi, G., and Dapas, B.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Cancers
• Publisher: MDPI
• ISSN: 2072-6694
• ISSN (Online): 2072-6694
• Published Online: 23 March 2022
• Copyright Holders: Copyright © 2022 The Authors
• First Published: First published in Cancers 14(7): 1630
• Publisher Policy: Reproduced under a Creative Commons License