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Kayeassamin A Isolated from the Flower of Mammea siamensis Triggers Apoptosis by Activating Caspase-3/-8 in HL-60 Human Leukemia Cells

1, 1, 2, 3, 1★★ Corresponding author

  1. 1Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki 859-3298, Japan
  2. 2Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002
  3. 3Faculty of Pharmacy, Payap University, Muang, Chiang Mai 50000, Thailand

CORRESPONDENCE

Yukihiro Shoyama

Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki 859-3298, Japan

shoyama@niu.ac.jp

Volume 8, Issue 4 · pp. 244-248 · 2016 · DOI: 10.4103/0974-8490.188884

ABSTRACT

Mammea siamensiŝ(Miq.) T. Anders. is used as a medicinal plant in Thailand and has several traditional therapeutic properties. In a previous study, we isolated eight compounds from the ˚ower of M. siamensis and demonstrated that kayeassamin A~(KA) exhibited potent antiproliferative activity against human leukemia and stomach cancer cell lines. In this study, we investigated the effect of KA on cell viability and apoptotic mechanisms in HL-60 human leukemia cells. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Nuclear morphology and DNA fragmentation were observed using Hoechst 33258 staining and agarose gel electrophoresis, respectively. The sub-G1 phase of cells was analyzed by ˚ow cytometry after the cellular DNA had been stained with propidium iodide. The protein levels of poly~ (ADP-ribose) polymerase~ (PARP) and caspases were determined by Western blotting. KA exhibited a signi˛cant cytotoxic effect in a dose-~ and time-dependent manner, and induced chromatin condensation, DNA fragmentation, and sub-G1 phase DNA content, known as molecular events associated with the induction of apoptosis. In addition, KA strongly induced the activation of PARP and caspase-3 and~-8, with weak caspase-9 activation. Furthermore, KA-induced DNA fragmentation was abolished by pretreatment with z-VAD-FMK (a broad caspase inhibitor), z-DEVD-FMK (a caspase-3 inhibitor), and z-IETD-FMK~ (a caspase-8 inhibitor), but not by z-LEHD-FMK (a caspase-9 inhibitor) pretreatment. These results indicate that KA triggers apoptotic cell death by activation of caspase-3 and~-8 in HL-60~cells.

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    Uto, T., Tung, N. H., Thongjankaew, P., Lhieochaiphant, S., & Shoyama, Y. (2016). Kayeassamin A Isolated from the Flower of Mammea siamensis Triggers Apoptosis by Activating Caspase-3/-8 in HL-60 Human Leukemia Cells. Pharmacognosy Research, 8(4), 244-248. https://doi.org/10.4103/0974-8490.188884

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    © 2016 Author(s)

    This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms. CC BY-NC-SA 4.0