NPC-14686 (Fmoc-l-homophenylalanine)-induced CaCa2+ increases and death in human prostate cancer cells.
The effect of NPC-14686, a potential anti-inflammatory drug, on cytosolic free Ca2+ levels ([Ca2+]i) and growth in PC3 human prostate cancer cells was examined by using fura-2 as a fluorescent Ca2+ indicator and WST-1 as a fluorescent growth dye. NPC-14686 at concentrations above 10 microM increased [Ca2+]i in a concentration-dependent manner with an EC50 value of 100 microM. NPC-14686-induced Ca2+ influx was confirmed by Mn2+ quench of fura-2 fluorescence. The Ca2+ signal was also reduced by removing extracellular Ca2+. Pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) to deplete the endoplasmic reticulum Ca2+ nearly abolished 200 microM NPC-14686-induced Ca2+ release; and conversely pretreatment with NPC-14686 completely inhibited thapsigargin-induced Ca2+ release. The Ca2+ release induced by 200 microM NPC-14686 was not affected by inhibiting phospholipase C with 2 microM U73122. Overnight treatment with 1-500 microM NPC-14686 decreased cell viability in a concentration-dependent manner. These findings suggest that in human PC3 prostate cancer cells, NPC-14686 increases [Ca2+]i by evoking extracellular Ca2+ influx and releasing intracellular Ca2+ from the endoplasmic reticulum via a phospholiase C-independent manner. NPC-14686 may be cytotoxic to prostate cancer cells.
Huang
Jong-Khing
JK
Department of Surgery, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan 813.
Liu
Chung-Shin
CS
Chen
Wei-Chuan
WC
Liu
Shiuh-Inn
SI
Hsu
Shu-Shong
SS
Chang
Hong-Tai
HT
Chou
Chiang-Ting
CT
Hsieh
Ching-Hong
CH
Chang
Chih-Hung
CH
Jan
Chung-Ren
CR
D000613:Aminobutyrates / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D005456:Fluorescent Dyes; D016257:Fura-2; D006801:Humans; D008297:Male; D008345:Manganese / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D013778:Tetrazolium Salts; D010738:Type C Phospholipases / Q000502:physiology
C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D000613:Aminobutyrates; D002121:Calcium Channel Blockers; D005456:Fluorescent Dyes; C417979:NPC 14686; D013778:Tetrazolium Salts; D008345:Manganese; D010738:Type C Phospholipases; D002118:Calcium; D016257:Fura-2