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Mechanism of apoptosis induced by diazoxide, a K+ channel opener, in HepG2 Human hepatoma cells

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Abstract

The effect of diazoxide, a K+ channel opener, on apoptotic cell death was investigated in HepG2 human hepatoblastoma cells. Diazoxide induced apoptosis in a dose-dependent manner and this was evaluated by flow cytometric assays of annexin-V binding and hypodiploid nuclei stained with propidium iodide. Diazoxide did not alter intracellular K+ concentration, and various inhibitors of K+ channels had no influence on the diazoxide-induced apoptosis; this implies that K+ channels activated by diazoxide may be absent in the HepG2 cells. However, diazoxide induced a rapid and sustained increase in intracellular Ca2+ concentration, and this was completely inhibited by the extracellular Ca2+ chelation with EGTA, but not by blockers of intracellular Ca2+ release (dantrolene and TMB-8). This result indicated that the diazoxideinduced increase of intracellular Ca2+ might be due to the activation of a Ca2+ influx pathway. Diazoxide-induced Ca2+ influx was not significantly inhibited by either voltage-operative Ca2+ channel blockers (nifedipine or verapamil), or by inhibitors of Na+, Ca2+-exchanger (bepridil and benzamil), but it was inhibited by flufenamic acid (FA), a Ca2+-permeable nonselective cation channel blocker. A quantitative analysis of apoptosis by flow cytometry revealed that a treatment with either FA or BAPTA, an intracellular Ca2+ chelator, significantly inhibited the diazoxide-induced apoptosis. Taken together, these results suggest that the observed diazoxideinduced apoptosis in the HepG2 cells may result from a Ca2+ influx through the activation of Ca2+-permeable non-selective cation channels. These results are very significant, and they lead us to further suggest that diazoxide may be valuable for the therapeutic intervention of human hepatomas.

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  1. College of Pharmacy, Duksung Women’s University, 132-714, Seoul, Korea

    Yong Soo Lee

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Lee, Y.S. Mechanism of apoptosis induced by diazoxide, a K+ channel opener, in HepG2 Human hepatoma cells. Arch Pharm Res 27, 305–313 (2004). https://doi.org/10.1007/BF02980065

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