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Effect of diindolylmethane on Ca2+ movement and viability in HA59T human hepatoma cells

  • Organ Toxicity and Mechanisms
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Abstract

The effect of diindolylmethane, a natural compound derived from indole-3-carbinol in cruciferous vegetables, on cytosolic Ca2+ concentrations ([Ca2+]i) and viability in HA59T human hepatoma cells is unclear. This study explored whether diindolylmethane changed [Ca2+]i in HA59T cells. The Ca2+-sensitive fluorescent dye fura-2 was applied to measure [Ca2+]i. Diindolylmethane at concentrations of 1–50 μM evoked a [Ca2+]i rise in a concentration-dependent manner. The signal was reduced by removing Ca2+. Diindolylmethane-induced Ca2+ influx was not inhibited by nifedipine, econazole, SK&F96365, and protein kinase C modulators but was inhibited by aristolochic acid. In Ca2+-free medium, treatment with the endoplasmic reticulum Ca2+ pump inhibitors thapsigargin or 2,5-di-tert-butylhydroquinone (BHQ) inhibited or abolished diindolylmethane-induced [Ca2+]i rise. Incubation with diindolylmethane inhibited thapsigargin or BHQ-induced [Ca2+]i rise. Inhibition of phospholipase C with U73122 reduced diindolylmethane-induced [Ca2+]i rise. At concentrations of 10–75 μM, diindolylmethane killed cells in a concentration-dependent manner. The cytotoxic effect of diindolylmethane was not reversed by chelating cytosolic Ca2+ with 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid. Propidium iodide staining data suggest that diindolylmethane (25–50 μM) induced apoptosis in a concentration-dependent manner. Collectively, in HA59T cells, diindolylmethane induced a [Ca2+]i rise by causing phospholipase C-dependent Ca2+ release from the endoplasmic reticulum and Ca2+ influx via phospholipase A2-sensitive channels. Diindolylmethane induced cell death that may involve apoptosis.

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Acknowledgments

This work was supported by grants from Kaohsiung Veterans General Hospital (VGHKS99-098) to C. R. Jan and VHYK-9901 to J. S. Cheng.

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Authors and Affiliations

  1. Department of Medicine, Yongkang Veterans Hospital, Tainan, 710, Taiwan

    Jin-Shiung Cheng & Kuo-Liang Chai

  2. Department of Surgery, Kaohsiung Veterans General Hospital, Kaohsiung, 813, Taiwan

    Su-Shung Shu

  3. Department of Nursing, Tzu Hui Institute of Technology, Pingtung, 926, Taiwan

    Chun-Chi Kuo

  4. Institute of Nursing and Department of Nursing, Chang Gung Institute of Technology, Chiayi Campus, Chiayi, 613, Taiwan

    Chiang-Ting Chou

  5. Department of Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, 813, Taiwan

    Wei-Lun Tsai

  6. Laboratory Medicine Division, Zuoying Armed Forces General Hospital, Kaohsiung, 813, Taiwan

    Yi-Chien Fang

  7. Department of Pathology and Laboratory Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, 813, Taiwan

    Li-Ni Kuo & Jeng-Hsien Yeh

  8. Department of Surgery, Ping Tung Christian Hospital, Ping Tung, 900, Taiwan

    Wei-Chuan Chen

  9. Department of Pediatrics, Ping Tung Christian Hospital, Ping Tung, 900, Taiwan

    Jau-Min Chien

  10. Department of Psychiatry, Kaohsiung Veterans General Hospital, Kaohsiung, 813, Taiwan

    Ti Lu & Chih-Chuan Pan

  11. Chang Bing Show Chwan Memorial Hospital, Changhua County, 505, Taiwan

    He-Hsiung Cheng

  12. Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung, 813, Taiwan

    Chung-Ren Jan

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  1. Jin-Shiung Cheng
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Correspondence to Chung-Ren Jan.

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Cheng, JS., Shu, SS., Kuo, CC. et al. Effect of diindolylmethane on Ca2+ movement and viability in HA59T human hepatoma cells. Arch Toxicol 85, 1257–1266 (2011). https://doi.org/10.1007/s00204-011-0670-9

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