Apoptosis induction preceded by mitochondrial depolarization in multiple myeloma cell line U266 by 2-aminophenoxazine-3-one

Ken Shirato, Kazuhiko Imaizumi, Keisuke Miyazawa, Akira Takasaki, Junichiro Mizuguchi, Xiao Fang Che, Shinichi Akiyama, Akio Tomoda

    Research output: Contribution to journalArticle

    15 Citations (Scopus)

    Abstract

    The aim of the present study was to investigate the mechanism of apoptosis in human multiple myeloma cell line, U266, caused by 2-aminophenoxazine-3-one (Phx-3). Flow-cytometrical and morphological analyses showed that Phx-3 increased the population of annexin V-positive cells including early stage apoptotic cells and late stage apoptotic cells and induced DNA fragmentation or apoptotic body formation in U266 cells, indicating that Phx-3 induced the apoptosis of U266 cells. Activity of caspase-3 was extensively increased in U266 cells treated with Phx-3 time-dependently within 24 h, but this Phx-3-stimulated activity of the enzyme in the cells was completely cancelled by the addition of N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk), a pan-caspase inhibitor. The addition of z-VAD-fmk almost blocked the apoptotic effect of Phx-3 against U266 cells, indicating that Phx-3-induced apoptosis of U266 cells was dependent on a caspase signaling pathway. Moreover, the apoptosis of U266 cells occurred after the induction of cell cycle arrest of the cells in the S and G2/M phase, the loss of mitochondrial membrane potential, and activation of caspase-3 reached maximum, which were caused by Phx-3 within 24 h. These results support the views that the apoptosis of U266 cells caused by Phx-3 may be preceded by the cell cycle arrest, depolarization of mitochondria and activation of caspase-3. These results support the view that Phx-3 may be utilized in future as chemotherapeutic agent against multiple myeloma which is extremely refractory to chemotherapy.

    Original languageEnglish
    Pages (from-to)62-67
    Number of pages6
    JournalBiological and Pharmaceutical Bulletin
    Volume31
    Issue number1
    DOIs
    Publication statusPublished - 2008 Jan

    Fingerprint

    Multiple Myeloma
    Apoptosis
    Cell Line
    Caspase 3
    Cell Cycle Checkpoints
    3-aminophenoxazone
    Caspase Inhibitors
    Mitochondrial Membrane Potential
    G2 Phase
    Annexin A5
    DNA Fragmentation
    Caspases
    Cell Division
    Mitochondria
    Drug Therapy

    Keywords

    • 2-aminophenoxazine-3-one
    • Apoptosis
    • Caspase-3
    • Mitochondrial membrane potential
    • Multiple myeloma

    ASJC Scopus subject areas

    • Molecular Medicine
    • Pharmacology, Toxicology and Pharmaceutics(all)

    Cite this

    Apoptosis induction preceded by mitochondrial depolarization in multiple myeloma cell line U266 by 2-aminophenoxazine-3-one. / Shirato, Ken; Imaizumi, Kazuhiko; Miyazawa, Keisuke; Takasaki, Akira; Mizuguchi, Junichiro; Che, Xiao Fang; Akiyama, Shinichi; Tomoda, Akio.

    In: Biological and Pharmaceutical Bulletin, Vol. 31, No. 1, 01.2008, p. 62-67.

    Research output: Contribution to journalArticle

    Shirato, K, Imaizumi, K, Miyazawa, K, Takasaki, A, Mizuguchi, J, Che, XF, Akiyama, S & Tomoda, A 2008, 'Apoptosis induction preceded by mitochondrial depolarization in multiple myeloma cell line U266 by 2-aminophenoxazine-3-one', Biological and Pharmaceutical Bulletin, vol. 31, no. 1, pp. 62-67. https://doi.org/10.1248/bpb.31.62
    Shirato, Ken ; Imaizumi, Kazuhiko ; Miyazawa, Keisuke ; Takasaki, Akira ; Mizuguchi, Junichiro ; Che, Xiao Fang ; Akiyama, Shinichi ; Tomoda, Akio. / Apoptosis induction preceded by mitochondrial depolarization in multiple myeloma cell line U266 by 2-aminophenoxazine-3-one. In: Biological and Pharmaceutical Bulletin. 2008 ; Vol. 31, No. 1. pp. 62-67.
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