Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0810, USA.
Biochem Pharmacol. 2011 Aug 15;82(4):418-25. doi: 10.1016/j.bcp.2011.05.013. Epub 2011 May 23.
G protein-coupled receptors, the largest cell surface receptor family, have emerged as critical players in cell death and survival. High gene expression level of the G(q)-coupled P2Y(1) nucleotide receptor in PC-3 prostate cancer cells was demonstrated using real-time quantitative PCR and confirmed by Western blotting and confocal laser scanning microscopy. A selective P2Y(1) receptor agonist, the ADP analogue MRS2365, concentration-dependently induced intracellular calcium mobilization (EC(50) 5.28nM), which was diminished by P2Y(1) receptor-selective antagonist MRS2500. P2Y(1) receptor activation by MRS2365 induced apoptosis in assays of Caspase-3, LDH release, and annexin-V staining. The pro-apoptotic effect of MRS2365 was blocked by MRS2500, P2Y(1) siRNA, and an inhibitor of the MAP kinase pathway PD98059. MRS2365 significantly inhibited the proliferation of PC-3 cells, examined using a MTT assay. Thus, activation of the P2Y(1) receptor induced cell death and inhibited growth of human prostatic carcinoma PC-3 cells. Activation of the P2Y(1) receptor should be a novel and promising therapeutic strategy for prostate cancer.
G 蛋白偶联受体是细胞表面受体家族中最大的家族，已成为细胞死亡和存活的关键参与者。使用实时定量 PCR 证明了 PC-3 前列腺癌细胞中 G(q)偶联 P2Y(1)核苷酸受体的高基因表达水平，并通过 Western blot 和共聚焦激光扫描显微镜得到证实。选择性 P2Y(1)受体激动剂，ADP 类似物 MRS2365，浓度依赖性地诱导细胞内钙动员（EC(50) 5.28nM），该作用被 P2Y(1)受体选择性拮抗剂 MRS2500 减弱。MRS2365 激活 P2Y(1)受体可诱导 Caspase-3、LDH 释放和 Annexin-V 染色检测到的细胞凋亡。MRS2365 的促凋亡作用被 MRS2500、P2Y(1)siRNA 和 MAP 激酶途径抑制剂 PD98059 阻断。MRS2365 显著抑制了 MTT 测定法检测到的 PC-3 细胞的增殖。因此，P2Y(1)受体的激活诱导了人前列腺癌细胞的死亡和抑制生长。激活 P2Y(1)受体应该是前列腺癌的一种新的有前途的治疗策略。
