resumo
The androgens/androgen receptor (AR) axis is the main therapeutic target in prostate cancer (PCa). However, while initially responsive, a subset of tumors loses AR expression through mechanisms putatively associated with epigenetic modifications. In this study, we assessed the link between the presence of CpG methylation in the 5 ' UTR and promoter regions of AR and loss of AR expression. Hence, we characterized and compared the methylation signature at CpG resolution of these regulatory regions in vitro, both at basal levels and following treatment with 5-aza-2-deoxycytidine (DAC) alone, or in combination with Trichostatin A (TSA). Our results showed heterogeneity in the methylation signature of AR negative cell lines and pinpointed the proximal promoter region as the most consistently methylated site in DU-145. Furthermore, this region was extremely resistant to the demethylating effects of DAC and was only significantly demethylated upon concomitant treatment with TSA. Nevertheless, no AR re-expression was detected at the mRNA or protein level. Importantly, after treatment, there was a significant increase in repressive histone marks at AR region 1 in DU-145 cells. Altogether, our data indicate that AR region 1 genomic availability is crucial for AR expression and that the inhibition of histone methyltransferases might hold promise for AR re-expression.
palavras-chave
DNA METHYLATION; GENE; TRANSCRIPTION; PROMOTER; SP1
categoria
Biochemistry & Molecular Biology
autores
Camilo, V; Pacheco, MB; Moreira-Silva, F; Outeiro-Pinho, G; Gaspar, VM; Mano, JF; Marques, CJ; Henrique, R; Jerónimo, C
nossos autores
João Mano
Vice-Diretor
Vítor Gaspar
Professor Auxiliaragradecimentos
The authors would like to acknowledge the Faculty of Medicine of the University of Porto for the support in conducting TOPO TA cloning and bacterial transformation.