Novel Effect of CP55,940, a CB1/CB2 Cannabinoid Receptor Agonist, on Intracellular Free Ca(superscript 2+) Levels in Bladder Cancer Cells

Chung-Ren Jan；Yih-Chau Lu；Bang-Ping Jiann；Hong-Tai Chang；Warren Su；Wei-Chung Chen；Jong-Khing Huang

CP55 ； 940 ； Ca(superscript 2+) signaling ； fura-2 ； BFTC cells ； bladder cancer

The Chinese Journal of Physiology

45卷1期（2002 / 03 / 01）

33 - 39

英文

The study was undertaken to explore the effect of CP55,940 ((-)-cis-3-[2-Hydroxy-4-(1,1-dimethylheptyl) pheuyl]-trans-4-(3-hydroxypropyl)cyclohexanol), a drug commonly used as a CB1/CB2 cannabinoid receptor agonist, on intracellular free Ca(superscript 2+) levels ([Ca(superscript 2+)](subscript i)) in several cell types. [Ca(superscript 2+)](subscript i) was measured in suspended cells by using the fluorescent dye fura-2 as an indicator. At concentrations between 1-50 μM, CPS5,940 increased [Ca(superscript 2+)](subscript i) in a concentration-dependent manner with an EC50 of 8 μM. The [Ca(superscript 2+)](subscript i) signal comprised an initial rise, a slow decay, and a sustained phase. CP55940 (10 μM)-induced [Ca(superscript 2+)](subscript i) signal was not altered by 5 μM of two cannabinoid receptor antagonists (AM-251, N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide; AM-281, 1-(2,4-Dichlorophenyl)-5-(4-iodophenyl)-4-m3thyl-N-4-morpholinyl-1H-pyrazole-3-carboxamide). Extracellular Ca(superscript 2+) removal decreased the maximum value of the Ca(superscript 2+) signals by 50%. CP55,940 (10 μM)-induced [Ca(superscript 2+)](subscript i) increase in Ca(superscript 2+)-free medium was inhibited by 80% by pretreatment with 1 μM thapsigargin, an endoplasmic reticulum Ca(superscript 2+) pump inhibitor. Conversely pretreatment with 10 μM CP55,940 in Ca(superscript 2+)-free medium for 6 min abolished thapsigargin-induced [Ca(superscript 2+)](subscript i) increase. Nifedipine (10 μM) and verapamil (10 μM) did not alter CP55,940 (10 μM)-induced [Ca(superscript 2+)](subscript i) increase. CP55, 940(10 μM)-iuduced Ca(superscript 2+) release was not affected when phospholipase C was inhibited by 2 μM U73122 (1-(6-((17β-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione). CP55,940 (5 μM) also increased [Ca(superscript 2+)](subscript i) in Madin-Darby canine kidney cells, MG63 human osteosarcoma cells, and IMR-32 neuroblastoma cells. Collectively, CP,55940 induced significant [Ca(superscript 2+)](subscript i) increases in several cell types by releasing store Ca2 from thapsigargin-sensitive pools and by causing Ca2 entry. The CPSS,940's action appears to be dissociated from stimulation of cannabinoid receptors.

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