Effect of Methoxsalen on Ca^(2+) Homeostasis and Viability in Human Osteosarcoma Cells

10.4077/CJP.2017.BAF482

Yi-Chau Lu；Chiang-Ting Chou；Wei-Zhe Liang；Chun-Chi Kuo；Shu-Shong Hsu；Jue-Long Wang；Chung-Ren Jan

Ca^(2+) ； endoplasmic reticulum ； human osteosarcoma cells ； methoxsalen ； phospholipase C

The Chinese Journal of Physiology

60卷3期（2017 / 06 / 30）

174 - 182

英文

Methoxsalen is a natural compound found in many seed plants. The effect of methoxsalen on Ca^(2+)-related physiology in human osteosarcoma is unclear. This study investigated the effect of methoxsalen on cytosolic free Ca^(2+) concentrations ([Ca^(2+)]_i) in MG63 human osteosarcoma cells. Methoxsalen induced [Ca^(2+)]_i rises concentration-dependently. Methoxsalen-induced Ca^(2+) entry was confirmed by Mn^(2+)-induced quench of fura-2 fluorescence. This Ca^(2+) entry was suppressed by nifedipine, econazole, and SKF96365. In Ca^(2+)-free medium, incubation with the endoplasmic reticulum Ca^(2+) pump inhibitor 2,5-di-tert-butylhydroquinone (BHQ) inhibited methoxsalen-evoked [Ca^(2+)]_i rises by 96%. In contrast, incubation with methoxsalen abolished BHQ-evoked [Ca^(2+)]_i rises. Inhibition of phospholipase C (PLC) with U73122 abolished methoxsalen-induced [Ca^(2+)]_i rises. Methoxsalen was cytotoxic at 300-700 μM in a concentration-dependent fashion. Chelating cytosolic Ca^(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxymethyl ester (BAPTA/AM) did not prevent methoxsalen-induced cytotoxicity. Collectively, our data suggest that in MG63 cells, methoxsalen induced [Ca^(2+)]_i rises by evoking PLC-dependent Ca^(2+) release from the endoplasmic reticulum, and Ca^(2+) entry via store-operated Ca^(2+) entry. Methoxsalen also induced Ca^(2+)-disassociated cell death.

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