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

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Abstract
Pharmacognosy Research,2016,8,4,244-248.
Published:August 2016
Type:Original Article
Authors:
Author(s) affiliations:

Takuhiro Uto1, Nguyen Huu Tung2, Pinjutha Thongjankaew3, Sorasak Lhieochaiphant4, Yukihiro Shoyama1

1Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki 859-3298, JAPAN.

2Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki 859-3298, Japan; School of Medicine and Pharmacy, Vietnam National University, Hanoi, Vietnam, THAILAND.

3Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand 4 Faculty of Pharmacy, Payap University, Muang, Chiang Mai 50000, THAILAND.

Abstract:

Background: Mammea siamensis (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 flower of M. siamensis and demonstrated that kayeassamin A (KA) exhibited potent antiproliferative activity against human leukemia and stomach cancer cell lines. Objective: In this study, we investigated the effect of KA on cell viability and apoptotic mechanisms in HL‑60 human leukemia cells. Materials and Methods: 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 flow 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. Results: KA exhibited a significant 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. Conclusion: These results indicate that KA triggers apoptotic cell death by activation of caspase‑3 and ‑8 in HL‑60 cells.

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Chemical structure of kayeassamin A

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