Effect of NPC15199 on [Ca^(2+)]_i and Viability in SCM1 Human Gastric Cancer Cells

10.4077/CJP.2016.BAF421

He-Hsiung Cheng；Chiang-Ting Chou；Wei-Zhe Liang；Jin-Shiung Cheng；Hong-Tai Chang；Chun-Chi Kuo；I-S Chen；Ti Lu；C-C Yu；Fu-An Chen；Daih-Huang Kuo；Po-Chuen Shieh；Chung-Ren Jan

Ca^(2+) ； endoplasmic reticulum ； fura-2 ； gastric cancer cells ； NPC15199

The Chinese Journal of Physiology

59卷5期（2016 / 10 / 31）

268 - 275

英文

NPC15199 is a synthesized compound that inhibits inflammation in some models. However, whether NPC15199 affects Ca^(2+) homeostasis in human gastric cancer is unclear. This study examined the effect of NPC15199 on cytosolic free Ca^(2+) concentrations ([Ca^(2+)]_i) and viability in SCM1 human gastric cancer cells. The Ca^(2+)-sensitive fluorescent dye fura-2 was used to measure [Ca^(2+)]_i. NPC15199 evoked [Ca^(2+)]_i rises concentration-dependently. The response was reduced by removing extracellular Ca^(2+). NPC15199-evoked Ca^(2+) entry was not inhibited by store-operated channel inhibitors (nifedipine, econazole and SKF96365) and protein kinase C (PKC) activator (phorbol 12-myristate 13 acetate, PMA), or PKC inhibitor (GF109203X). In Ca^(2+)-free medium, treatment with the endoplasmic reticulum Ca^(2+) pump inhibitor thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) nearly abolished NPC15199-evoked [Ca^(2+)]_i rises. Conversely, treatment with NPC15199 also nearly abolished thapsigargin or BHQ-evoked [Ca^(2+)]_i rises. Inhibition of phospholipase C (PLC) with U73122 did not affect NPC15199-evoked [Ca^(2+)]_i rises. NPC15199 at concentrations of 100-900 μM induced concentration-dependent, Ca^(2+)-independent decrease in viability. Together, in SCM1 cells, NPC15199 induced [Ca^(2+)]_i rises that involved Ca^(2+) entry through PKC-insensitive non-store-operated Ca^(2+) channels and PLC-independent Ca^(2+) release from the endoplasmic reticulum. NPC15199 also induced Ca^(2+)-independent cell death.

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