Shabbir Majid, Ryten Mina, Thompson Cecil, Mikhailidis Dimitri, Burnstock Geoffrey
Autonomic Neuroscience Centre, Royal Free and University College Medical School, London, UK.
BJU Int. 2008 Feb;101(3):352-9. doi: 10.1111/j.1464-410X.2007.07293.x. Epub 2007 Nov 13.
To investigate the nature of purinergic signalling in hormone-refractory prostate cancer (HRPC) cells in vitro, as extracellular ATP inhibits the growth of HRPC in vitro via the activation of P2 purinergic receptors, and to characterize which P2 receptors subtypes and secondary mechanisms are involved.
The effect of extracellular ATP on HRPC cell lines PC-3 and DU-145, and the normal prostate cell line PNT-2, were investigated. Reverse-transcription polymerase chain reaction was used to assess P2 purinergic receptors, which were pharmacologically characterized using various receptor agonists and antagonists. The effect of ATP on intracellular Ca(2+) concentration (Ca(2+)) was examined to asses its role in growth inhibition. The effect of combining ATP with the chemotherapeutic drug mitoxantrone was also assessed.
PC-3 cells expressed mRNA for P2X(4,5,7), P2Y(1,2,4,6); DU-145 cells expressed mRNA for P2X(4,5), P2Y(1,2,4,6,11); PNT-2 cells expressed mRNA for P2X(4,5,7) and P2Y(1,2,4,6,11). ATP (10(-4)m) inhibited HRPC PC-3 cell growth by approximately 90%, an effect partially inhibited by the nonselective P2 receptor antagonists pyridoxal-5'-phosphate-6-azophenyl-2',4' disulphonic acid (PPADS) and suramin. The order of potency of agonists was: adenosine 5'-O-(3 thiotriphosphate) > ATP > benzoyl benzoyl ATP >> 2-methylthio ATP. DU-145 cells responded similarly. Pharmacological profiling implicated P2X(5) and/or P2Y(11) receptors in the antineoplastic response in HRPC. ATP induced apoptosis in a Ca(2+)-independent mechanism. ATP was significantly less effective on PNT-2 cells, which also had a different order of agonist potency. ATP combined with mitoxantrone in an additive manner in HRPC.
ATP effectively reduces growth of HRPC cells via calcium-independent apoptosis. Pharmacological profiling indicates P2X(5) and/or P2Y(11) receptors in this process, with a different functional purinergic receptor profile and sensitivity in normal vs cancer cells.
研究激素难治性前列腺癌（HRPC）细胞中嘌呤能信号传导的性质，因为细胞外ATP通过激活P2嘌呤能受体在体外抑制HRPC的生长，并确定涉及哪些P2受体亚型和次要机制。
研究细胞外ATP对HRPC细胞系PC-3和DU-145以及正常前列腺细胞系PNT-2的影响。采用逆转录聚合酶链反应评估P2嘌呤能受体，并用各种受体激动剂和拮抗剂对其进行药理学特性分析。检测ATP对细胞内Ca(2+)浓度(Ca(2+))的影响，以评估其在生长抑制中的作用。还评估了ATP与化疗药物米托蒽醌联合使用的效果。
PC-3细胞表达P2X(4,5,7)、P2Y(1,2,4,6)的mRNA；DU-145细胞表达P2X(4,5)、P2Y(1,2,4,6,11)的mRNA；PNT-2细胞表达P2X(4,5,7)和P2Y(1,2,4,6,11)的mRNA。ATP（10(-4)m）抑制HRPC PC-3细胞生长约90%，非选择性P2受体拮抗剂吡哆醛-5'-磷酸-6-偶氮苯-2',4'二磺酸（PPADS）和苏拉明部分抑制了该作用。激动剂的效力顺序为：腺苷5'-O-(3-硫代三磷酸) > ATP > 苯甲酰苯甲酰ATP >> 2-甲硫基ATP。DU-145细胞有类似反应。药理学分析表明P2X(5)和/或P2Y(11)受体参与了HRPC的抗肿瘤反应。ATP通过Ca(2+)非依赖机制诱导细胞凋亡。ATP对PNT-2细胞的作用明显较弱，其激动剂效力顺序也不同。在HRPC中，ATP与米托蒽醌以相加方式联合作用。
ATP通过钙非依赖凋亡有效降低HRPC细胞的生长。药理学分析表明在此过程中涉及P2X(5)和/或P2Y(11)受体，正常细胞与癌细胞的嘌呤能受体功能谱和敏感性不同。
