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3,3′‐Diindolylmethane Alters Ca 2+ Homeostasis and Viability in MG63 Human Osteosarcoma Cells
Author(s) -
Lu YiChau,
Chen IShu,
Chou ChiangTing,
Huang JongKhing,
Chang HongTai,
Tsai JengYu,
Hsu ShuShong,
Liao WeiChuan,
Wang JueLong,
Lin KoLong,
Liu ShuihInn,
Kuo ChunChi,
Ho ChinMan,
Jan ChungRen
Publication year - 2012
Publication title -
basic and clinical pharmacology and toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.805
H-Index - 90
eISSN - 1742-7843
pISSN - 1742-7835
DOI - 10.1111/j.1742-7843.2011.00816.x
Subject(s) - thapsigargin , endoplasmic reticulum , phospholipase c , fura 2 , bapta , annexin , propidium iodide , extracellular , ionomycin , microbiology and biotechnology , viability assay , chemistry , apoptosis , kinase , cytosol , biology , biochemistry , signal transduction , programmed cell death , intracellular , enzyme
The effect of the natural product 3,3′‐diindolylmethane (DIM) on cytosolic Ca 2+ concentrations ([Ca 2+ ] i ) and viability in MG63 human osteosarcoma cells was explored. The Ca 2+ ‐sensitive fluorescent dye fura‐2 was applied to measure [Ca 2+ ] i . DIM at concentrations of 40–80 μM induced a [Ca 2+ ] i rise in a concentration‐dependent manner. The response was reduced partly by removing Ca 2+ . DIM‐evoked Ca 2+ entry was suppressed by nifedipine, econazole, SK&F96365 and protein kinase C modulators. In the absence of extracellular Ca 2+ , incubation with the endoplasmic reticulum Ca 2+ pump inhibitors thapsigargin or 2,5‐di‐tert‐butylhydroquinone (BHQ) inhibited or abolished DIM‐induced [Ca 2+ ] i rise. Incubation with DIM also inhibited thapsigargin or BHQ‐induced [Ca 2+ ] i rise. Inhibition of phospholipase C with U73122 abolished DIM‐induced [Ca 2+ ] i rise. At concentrations of 10–50 μM, DIM killed cells in a concentration‐dependent manner. This cytotoxic effect was not altered by chelating cytosolic Ca 2+ with 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid (BAPTA). Annexin V/propidium iodide staining data implicate that DIM (20 and 40 μM) induced apoptosis in a concentration‐dependent manner. In sum, in MG63 cells, DIM induced a [Ca 2+ ] i rise by evoking phospholipase C‐dependent Ca 2+ release from the endoplasmic reticulum and Ca 2+ entry via protein kinase C‐sensitive store‐operated Ca 2+ channels. DIM caused cell death that may involve apoptosis.