PUBLICATION
5-Azacytidine Downregulates the Proliferation and Migration of Hepatocellular Carcinoma Cells In Vitro and In Vivo by Targeting miR-139-5p/ROCK2 Pathway
- 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., Dapas, B.
- ID
- ZDB-PUB-220413-2
- Date
- 2022
- Source
- Cancers 14(7): (Journal)
- Registered Authors
- Keywords
- 5-azacytidine, E2F1, MMP-2, ROCK2, cell cycle, hepatocellular carcinoma, miR-139-5p, migration
- MeSH Terms
- none
- PubMed
- 35406401 Full text @ Cancers
Citation
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., Dapas, B. (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):.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping