The Antidepressant Mirtazapine-Induced Cytosolic Ca(superscript 2+) Elevation and Cytotoxicity in Human Osteosarcoma Cells

Chin-Chuan Pan；He-Hsiung Cheng；Chun-Jen Huang；Yih-Chau Lu；I-Shu Chen；Shiuh-Inn Liu；Shu-Shong Hsu；Hong-Tai Chang；Jong-Khing Huang；Jin-Shyr Chen；Chung-Ren Jan

Ca(superscript 2+) ； fura-2 ； MG63 cells ； mirtazapine ； osteosarcoma cells ； thapsigargin

The Chinese Journal of Physiology

49卷6期（2006 / 12 / 01）

290 - 297

英文

The effect of the antidepressant mirtazapine on cytosolic free Ca(superscript 2+) concentration ([Ca(superscript 2+)](subscript i)) and viability has not been explored in any cell type. This study examined whether mirtazapine alters Ca(superscript 2+) levels and causes cell death in osteoblast-like cells using MG63 human osteosarcoma cells as a model. [Ca(superscript 2+)](subscript i) and cell viability were measured using the fluorescent dyes fura-2 and WST-1, respectively. Mirtazapine at concentrations above 250 μM increased [Ca(superscript 2+)](subscript i) in a concentration-dependent manner. The Ca(superscript 2+) signal was reduced by 60% by removing extracellular Ca(superscript 2+). The mirtazapine-induced Ca(superscript 2+) influx was sensitive to blockade of nifedipine and verapamil. In Ca(superscript 2+)-free medium, after pretreatment with 1.5 mM mirtazapine, 1 μM thapsigargin (an endoplasmic reticulum Ca(superscript 2+) pump inhibitor), 2 μM CCCP (a mitochondrial uncoupler), and 1 μM ionomycin failed to release more stored Ca(superscript 2+); conversely, pretreatment with thapsigargin, CCCP and ionomycin abolished mirtazapine-induced Ca(superscript 2+) release. Inhibition of phospholipase C with 2 μM U73122 did not change mirtazapine-induced [Ca(superscript 2+)](subscript i) increase. Seal of Ca(superscript 2+) movement across the plasma membrane with 50 μM extracellular La(superscript 3+) enhanced 1 μM thapsigargin-induced [Ca(superscript 2+)](subscript i) increase, suggesting that Ca(superscript 2+) efflux played a role in lowering thapsigargin-induced [Ca(superscript 2+)](subscript i) increase; however, the same La(superscript 3+) treatment did not alter mirtazapine-induced [Ca(superscript 2+)](subscript i) increase. At concentrations of 500 μM and 1000 μM, mirtazapine killed 30% and 60% cells, respectively. The cytotoxicity was not reversed by chelating cytosolic Ca(superscript 2+) with BAPTA. Collectively, in MG63 cells, mirtazapine induced a [Ca(superscript 2+)](subscript i) increase by causing Ca(superscript 2+) release from stores and Ca(superscript 2+) influx from extracellular space. Furthermore, mirtazapine caused cytotoxicity at higher concentrations in a Ca(superscript 2+)-dissociated manner.

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