resumo
Castration-resistant prostate cancer (CRPC) is an incurable form of prostate cancer (PCa), with DNMT1 and G9a being reported as overexpressed, rendering them highly attractive targets for precision medicine. CM-272 is a dual inhibitor of both methyltransferases' activity. Herein, we assessed the response of different PCa cell lines to CM-272, in both 2D and 3D models, and explored the molecular mechanisms underlying CM-272 inhibitory effects.CRPC tissues displayed significantly higher DNMT1, G9a and H3K9me2 expression than localized PCa. In vitro, CM-272 caused a significant decrease in PCa cell viability and proliferation alongside with increased apoptotic levels. We disclose that, under the evaluated dose, CM-272 led to G9a activity inhibition, while not significantly affecting DNMT1 activity. Upon G9a knockdown, DU145 and PC3 showed decreased cell viability. Remarkably, DU145 cells treated with CM-272 or with G9a knockdown displayed no differences in viability, suggesting a SET dependent mechanism. Contrarily, PC3 cell viability impact was higher in G9a knockdown, compared with CM 272 treatment, suggesting an additional G9a function. Moreover, DU145 cells overexpressing catalytically functional G9a disclosed higher resistance to CM-272 treatment, reinforcing that the drug mechanism of action is dependent on G9a catalytic function.Importantly, we successfully assembled spheroids from several prostate cell lines. Our results showed that CM 272 retained its anti-tumoral effects in 3D PCa models, leading to a clear reduction in cancer cell survival. We concluded that inhibition of G9a methyltransferase activity by CM-272 has anti-tumor effect in PCa cells, holding therapeutic potential against CRPC.
palavras-chave
HISTONE METHYLTRANSFERASE; DNA METHYLATION; H3K9; EXPRESSION; COMPLEXES; GENES
categoria
Research & Experimental Medicine; Pharmacology & Pharmacy
autores
Moreira-Silva, F; Outeiro-Pinho, G; Lobo, J; Guimaraes, R; Gaspar, VM; Mano, JF; Agirre, X; Pineda-Lucena, A; Prosper, F; Paramio, JM; Henrique, R; Correia, MP; Jeronimo, C
nossos autores
João Mano
Vice-Diretor
Vítor Gaspar
Professor Auxiliaragradecimentos
C.J. research is supported by Programa Operacional Competitividade e Internacionalizac ao (POCI), in the component FEDER, and by national funds (OE) through FCT/MCTES, in the scope of the project HyTherCaP- POCI-01-0145-FEDER-29030 (PTDC/MECONC/29030/2017) and by the Research Center of Portuguese Oncology Institute of Porto (CI-IPOP- 272016). V.M.G. and M.P.C. were funded by FCT-Fundacao para a Ciencia e Tecnologia (CEEC/1048/2019 and CEECINST/00091/2018, respectively). J.M.P. lab is partially funded by grants from the EuropeanRegional Development Fund (FEDER) for Science and Innovation (SAF2015-66015-R, and PID2019-110758RB-I00 and RED2018- 102769-T), and Instituto de Salud Carlos III (CIBERONC no. CB16/12/ 00228). A.P.-L. lab is partially funded by grants from the Spanish Ministry of Science and Innovation (PID2020- 115875RB-I00) and Fundacion Fuentes Dutor. F.M.-S. and G.O.-P. were funded by Porto Comprehensive Cancer Center (Porto.CCC, Contract RNCCCP.CCC-CI- IPOP-LAB3- NORTE-01-0145-FEDER-072678).