Department of Biotechnology, Institute of Graduate Studies and Research, Alexandria University, Alexandria, Egypt.
Department of Pharmacology & Toxicology, Faculty of Pharmacy, Damanhur University, Damanhur, Egypt.
Med Oncol. 2022 Jul 14;39(10):143. doi: 10.1007/s12032-022-01718-8.
Aberrant activation of several signaling pathways has been implicated in prostate cancer (PCa) progression to castrate-resistant prostate cancer (CRPC). Phosphoinositide-3-kinase/Protein Kinase B/mechanistic Target of Rapamycin (PI3K/AKT/mTOR) and Hedgehog/GLI (Hh/GLI) pathways are major participants in progression to CRPC. In this sense, the current work aims to assess the potential antitumor effects resulting from co-targeting the aforementioned pathways in PC3 cells with Dactolisib as a dual PI3K/mTOR inhibitor and GANT61 as a GLI1 antagonist. Three replica of PC3 cells were assigned for four treatment groups; vehicle control, Dactolisib-treated, GANT61-treated, and combination-treated groups. GLI1 gene expression was determined by quantitative real-time PCR while active caspase-3 was determined colorimetrically. P-AKT, p70 ribosomal s6 protein kinase 1 (pS6K1), cyclin D1, vascular endothelial growth factor 1 (VEGF1), and Microtubule-associated proteins 1A/1B light chain 3 (LC3) protein levels were determined by ELISA technique. GLI1 gene expression was down-regulated as a result of Dactolisib, GANT61, and their combination. Additionally, both drugs significantly reduced p-AKT, pS6K1, cyclin D1, and VEGF1 protein levels. Dactolisib elevated LC3 protein levels and GANT61 augmented Dactolisib effect on LC3. Moreover, only Dactolisib/GANT61combination significantly increased active caspase-3 level. To sum up, Dactolisib/GANT61 combination was shown to be promising in PCa treatment. Further in-vitro and in-vivo studies are warranted to support our findings.
几种信号通路的异常激活被认为与前列腺癌（PCa）向去势抵抗性前列腺癌（CRPC）的进展有关。磷脂酰肌醇 3-激酶/蛋白激酶 B/雷帕霉素（PI3K/AKT/mTOR）和 Hedgehog/GLI（Hh/GLI）途径是向 CRPC 进展的主要参与者。从这个意义上说，目前的工作旨在评估用 Dactolisib（一种双重 PI3K/mTOR 抑制剂）和 GANT61（一种 GLI1 拮抗剂）靶向上述途径对 PC3 细胞的潜在抗肿瘤作用。将三个 PC3 细胞复制品分配到四个治疗组：载体对照组、Dactolisib 处理组、GANT61 处理组和联合处理组。通过定量实时 PCR 确定 GLI1 基因表达，通过比色法确定活性 caspase-3。通过 ELISA 技术测定磷酸化 AKT（p-AKT）、p70 核糖体 S6 蛋白激酶 1（pS6K1）、细胞周期蛋白 D1、血管内皮生长因子 1（VEGF1）和微管相关蛋白 1A/1B 轻链 3（LC3）蛋白水平。Dactolisib、GANT61 及其组合均可下调 GLI1 基因表达。此外，两种药物均显著降低了 p-AKT、pS6K1、细胞周期蛋白 D1 和 VEGF1 蛋白水平。Dactolisib 增加了 LC3 蛋白水平，GANT61 增强了 Dactolisib 对 LC3 的作用。此外，只有 Dactolisib/GANT61 联合用药显著增加了活性 caspase-3 水平。总之，Dactolisib/GANT61 联合用药在前列腺癌治疗中显示出很大的潜力。需要进一步的体外和体内研究来支持我们的发现。
