The mechanism of NPC-14686-induced [Ca²⁺]i rises and non-Ca²⁺-triggered cell death in MG63 human osteosarcoma cells.

Abstract

NPC-14686 has been shown to have anti-inflammatory effect in previous studies, but the mechanisms are unclear. The effect of NPC-14686 on cytosolic Ca²⁺ concentrations ([Ca²⁺]i) and viability in MG63 human osteosarcoma cells was explored. The Ca²⁺-sensitive fluorescent dye fura-2 was applied to measure [Ca²⁺]i. NPC-14686 at concentrations of 100-500 μM induced a [Ca²⁺]i rise in a concentration-dependent manner. The response was reduced by 80% by removing Ca²⁺. NPC-14686 induced Mn²⁺ influx leading to quenching of fura-2 fluorescence. NPC-14686-evoked Ca²⁺ entry was suppressed by nifedipine, econazole, SK&F96365, and protein kinase C inhibitor. Inhibition of phospholipase C with U73122 abolished NPC-14686-induced [Ca²⁺]i rise. At 20-50 μM, NPC-14686 decreased cell viability, which was not reversed by chelating cytosolic Ca²⁺ with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl (BAPTA/AM). Annexin V/propidium iodide staining data suggest that NPC-14686 (30-50 μM) induced apoptosis in a concentration-dependent manner. NPC-14686 also increased levels of reactive oxygen species. Together, in human osteosarcoma cells, NPC-14686 induced a [Ca²⁺]i rise by inducing phospholipase C-dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via protein kinase C-sensitive store-operated Ca²⁺ channels. NPC-14686 induced cell death that might involve apoptosis via mitochondrial pathways.

• Authors: Jau-Min Chien, Chiang-Ting Chou, Wei-Zhe Liang, Daih-Huang Kuo, Chun-Chi Kuo, Chin-Man Ho, Pochuen Shieh, Chung-Ren Jan
• Journal: The Chinese journal of physiology (Chin J Physiol) Vol. 57 Issue 3 Pg. 158-66 (Jun 30 2014) ISSN: 0304-4920 [Print] India
• PMID: 24826784 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Aminobutyrates
• Antifungal Agents
• Calcium Channel Blockers
• Chelating Agents
• Imidazoles
• NPC 14686
• 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
• Egtazic Acid
• Econazole
• 1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole
• Nifedipine
• Calcium

Topics

• Aminobutyrates (pharmacology)
• Antifungal Agents (pharmacology)
• Apoptosis (drug effects, physiology)
• Bone Neoplasms (pathology)
• Calcium (metabolism)
• Calcium Channel Blockers (pharmacology)
• Calcium Signaling (drug effects, physiology)
• Cell Line, Tumor
• Cell Survival (drug effects, physiology)
• Chelating Agents (pharmacology)
• Econazole (pharmacology)
• Egtazic Acid (analogs & derivatives, pharmacology)
• Humans
• Imidazoles (pharmacology)
• Nifedipine (pharmacology)
• Osteosarcoma (pathology)