Effect of Methoxychlor on Ca^(2+) Homeostasis and Apoptosis in HA59T Human Hepatoma Cells

10.4077/CJP.2015.BAD276

Chi-Ting Horng；Chiang-Ting Chou；Hui-Wen Tseng；Jin-Shiung Cheng；Hong-Tai Chang；Po-Min Chang；I-Li Chen；Ming-Chi Hung；Yi-Jen Tsai；Peng-Chih Tsai；Wei-Zhe Liang；Chun-Chi Kuo；Daih-Huang Kuo；Chin-Man Ho；Jia-Rong Lin；Pochuen Shieh；Chung-Ren Jan

apoptosis ； Ca^(2+) ； hepatoma ； methoxychlor

The Chinese Journal of Physiology

58卷1期（2015 / 02 / 01）

1 - 8

英文

Methoxychlor, an organochlorine pesticide, is thought to be an endocrine disrupter that affects Ca^(2+) homeostasis and cell viability in different cell models. This study explored the action of methoxychlor on cytosolic free Ca^(2+) concentrations ([Ca^(2+)] i) and apoptosis in HA59T human hepatoma cells. Fura-2, a Ca^(2+)-sensitive fluorescent dye, was applied to measure [Ca^(2+)] i. Methoxychlor at concentrations of 0.1-1μM caused a [Ca^(2+)] i rise in a concentration-dependent manner. Removal of external Ca^(2+) abolished methoxychlor's effect. Methoxychlor-induced Ca^(2+) influx was confirmed by Mn^(2+)-induced quench of fura-2 fluorescence. Methoxychlor-induced Ca^(2+) entry was inhibited by nifedipine, econazole, SK&F96365, and protein kinase C modulators. Methoxychlor killed cells at concentrations of 10-130μM in a concentration-dependent fashion. Chelating of cytosolic Ca^(2+) with 1, 2-bis (2-aminophenoxy) ethane-N, N, N', N'-tetraacetic acid/AM (BAPTA/AM) did not prevent methoxychlor's cytotoxicity. Methoxychlor (10 and 50μM) induced apoptosis concentration-dependently as determined by using Annexin V/propidium iodide staining. Together, in HA59T cells, methoxychlor induced a [Ca^(2+)] i rise by inducing Ca^(2+) entry via protein kinase C-sensitive Ca^(2+)-permeable channels, without causing Ca^(2+) release from stores. Methoxychlor also induced apoptosis that was independent of [Ca^(2+)] i rises.

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