Cancer Biology and Epigenetics Group, Research Center of IPO Porto (CI-IPOP)/RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO Porto)/Porto Comprehensive Cancer Center (Porto.CCC) Raquel Seruca, R. Dr. António Bernardino de Almeida, 4200-072 Porto, Portugal.
CICECO-Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal.
Biomolecules. 2023 Oct 14;13(10):1526. doi: 10.3390/biom13101526.
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.
雄激素/雄激素受体(AR)轴是前列腺癌(PCa)的主要治疗靶点。然而,尽管最初有反应,但一部分肿瘤通过与表观遗传修饰相关的机制失去 AR 表达。在这项研究中,我们评估了 AR 的 5'UTR 和启动子区域的 CpG 甲基化与 AR 表达缺失之间的联系。因此,我们在体外以 CpG 分辨率表征和比较了这些调节区域的甲基化特征,包括基础水平以及单独用 5-氮杂-2-脱氧胞苷(DAC)或与 Trichostatin A(TSA)联合处理后的水平。我们的结果显示 AR 阴性细胞系的甲基化特征存在异质性,并指出近端启动子区域是 DU-145 中最一致甲基化的位点。此外,该区域对 DAC 的去甲基化作用具有极强的抗性,仅在与 TSA 联合处理时才会显著去甲基化。然而,在 mRNA 或蛋白水平均未检测到 AR 的重新表达。重要的是,治疗后,在 DU-145 细胞中,AR 区域 1 的抑制性组蛋白标记显著增加。总之,我们的数据表明 AR 区域 1 的基因组可用性对于 AR 表达至关重要,抑制组蛋白甲基转移酶可能为 AR 的重新表达带来希望。