2. Academic Validation
  3. Effect of methoxychlor on Ca²⁺ homeostasis and apoptosis in HA59T human hepatoma cells

Effect of methoxychlor on Ca²⁺ homeostasis and apoptosis in HA59T human hepatoma cells

  • Chin J Physiol. 2015 Feb 28;58(1):1-8. doi: 10.4077/CJP.2015.BAD276.
Chi-Ting Horng 1,  Chiang-Ting Chou 2,  Hui-Wen Tseng 3,  Jin-Shiung Cheng 4,  Hong-Tai Chang 5,  Po-Min Chang 5,  I-Li Chen 6,  Ming-Chi Hung 6,  Yi-Jen Tsai 6,  Peng-Chih Tsai 6,  Wei-Zhe Liang 7,  Chun-Chi Kuo 8,  Daih-Huang Kuo 6,  Chin-Man Ho 7,  Jia-Rong Lin 7,  Pochuen Shieh 6,  Chung-Ren Jan 7
Affiliations

Affiliations

  • 1 Department of Ophthalmology, Kaohsiung Armed Force General Hospital, Kaohsiung 80284, Taiwan, Republic of China.
  • 2 Department of Nursing, Division of Basic Medical Sciences, Chang Gung University of Science and Technology, Chia-Yi 61363, Taiwan, Republic of China.
  • 3 Department of Dermatology, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan, Republic of China.
  • 4 Department of Medicine, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan, Republic of China.
  • 5 Department of Surgery, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan, Republic of China.
  • 6 Department of Pharmacy, Tajen University, Pingtung 90741, Taiwan, Republic of China.
  • 7 Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan, Republic of China.
  • 8 Department of Nursing, Tzu Hui Institute of Technology, Pingtung 92641, Taiwan, Republic of China.
PMID: 25687486 DOI: 10.4077/CJP.2015.BAD276
Abstract

Methoxychlor, an organochlorine pesticide, is thought to be an endocrine disrupter that affects Ca²⁺ homeostasis and cell viability in different cell models. This study explored the action of methoxychlor on cytosolic free Ca²⁺ concentrations ([Ca²⁺]i) and Apoptosis in HA59T human hepatoma cells. Fura-2, a Ca²⁺-sensitive Fluorescent Dye, was applied to measure [Ca²⁺]i. Methoxychlor at concentrations of 0.1-1 μM caused a [Ca²⁺]i rise in a concentration-dependent manner. Removal of external Ca²⁺ abolished methoxychlor's effect. Methoxychlor-induced Ca²⁺ influx was confirmed by Mn²⁺-induced quench of fura-2 fluorescence. Methoxychlor-induced Ca²⁺ entry was inhibited by nifedipine, econazole, SK&F96365, and protein kinase C modulators. Methoxychlor killed cells at concentrations of 10-130 μM in a concentration-dependent fashion. Chelation of cytosolic Ca²⁺ 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 50 μM) induced Apoptosis concentration-dependently as determined by using Annexin V/propidium iodide staining. Together, in HA59T cells, methoxychlor induced a [Ca²⁺]i rise by inducing Ca²⁺ entry via protein kinase C-sensitive Ca²⁺-permeable channels, without causing Ca²⁺ release from stores. Methoxychlor also induced Apoptosis that was independent of [Ca²⁺]i rises.

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