Kayeassamin A Isolated from the Flower ofMammea siamensisTriggers Apoptosis by Activating Caspase-3/-8 in HL-60 Human Leukemia Cells.
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.


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.


CONCLUSIONS
These results indicate that KA triggers apoptotic cell death by activation of caspase-3 and -8 in HL-60 cells.


CONCLUSIONS
Kayeassamin A (KA) isolated from the flower ofMammea siamensisexhibited a significant cytotoxic effect in HL-60 human leukemia cells. KA triggers apoptotic cell death by activating caspase-3/-8.Abbreviations Used: KA: Kayeassamin A; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PARP: Poly (ADP-ribose) polymerase; PI: Propidium iodide; CA: Corosolic acid.
Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki 859-3298, Japan.
Takuhiro
T
Uto
Department 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.
Nguyen Huu
NH
Tung
Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
Pinjutha
P
Thongjankaew
Faculty of Pharmacy, Payap University, Muang, Chiang Mai 50000, Thailand.
Sorasak
S
Lhieochaiphant
Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki 859-3298, Japan.
Yukihiro
Y
Shoyama
Apoptosis; Mammea siamensis; caspase; kayeassamin A; leukemia
Results Probl Cell Differ. 1999;23:25-63
9950028
Am J Chin Med. 2015;43(4):757-67
26119959
Phytother Res. 2015 Dec;29(12 ):1956-63
26463105
Phytochemistry. 1998 Apr;47(8):1661-3
9612959
Anticancer Agents Med Chem. 2008 Apr;8(3):322-41
18393791
Phytochemistry. 2006 May;67(9):924-8
16563448
Bioorg Med Chem Lett. 2013 Jan 1;23(1):158-62
23206866
Radiat Res. 1998 Jul;150(1):3-10
9650595
Bioorg Med Chem. 2012 Aug 15;20(16):4968-77
22831798
Bioorg Med Chem. 2008 Sep 15;16(18):8653-60
18725180
Exp Oncol. 2007 Sep;29(3):175-80
18004240
Int J Mol Sci. 2013 Feb 18;14(2):4106-20
23429195
Cell Biochem Funct. 2011 Aug;29(6):468-80
21773978
Curr Med Chem. 2005;12(8):887-916
15853704
J Nat Prod. 2002 May;65(5):757-60
12027761
Biol Chem. 2013 Jul;394(7):831-43
23509217
Anticancer Drugs. 2013 Nov;24(10):1058-68
24025561
Toxicol Pathol. 2007 Jun;35(4):495-516
17562483
Oncogene. 2003 Nov 24;22(53):8543-67
14634618
Bioorg Med Chem Lett. 2008 Aug 15;18(16):4688-91
18640837