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Sulforaphane‐induced apoptosis in human leukemia H L ‐60 cells through extrinsic and intrinsic signal pathways and altering associated genes expression assayed by c DNA microarray
Author(s) -
Shang HungSheng,
Shih YungLuen,
Lee ChingHsiao,
Hsueh ShuChing,
Liu JiaYou,
Liao NienChieh,
Chen YungLiang,
Huang YiPing,
Lu HsuFeng,
Chung JingGung
Publication year - 2017
Publication title -
environmental toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 77
eISSN - 1522-7278
pISSN - 1520-4081
DOI - 10.1002/tox.22237
Subject(s) - apoptosis , sulforaphane , xiap , microbiology and biotechnology , cell cycle , chemistry , viability assay , fas ligand , biology , fadd , caspase , programmed cell death , biochemistry
Sulforaphane (SFN), one of the isothiocyanates, is a biologically active compound extracted from cruciferous vegetables, and has been shown to induce cytotoxic effects on many human cancer cells including human leukemia cells. However, the exact molecular mechanism and altered gene expression associated with apoptosis is unclear. In this study, we investigated SFN‐induced cytotoxic effects and whether or not they went through cell‐cycle arrest and induction of apoptosis and further examined molecular mechanism and altered gene expression in human leukemia HL‐60 cells. Cell viability, cell‐cycle distribution, sub‐G1 (apoptosis), reactive oxygen species (ROS) and Ca 2+ production, levels of mitochondrial membrane potential (ΔΨ m ), and caspase‐3, −8, and −9 activities were assayed by flow cytometry. Apoptosis‐associated proteins levels and gene expressions were examined by Western blotting and cDNA microarray assays, respectively. Results indicated that SFN decreased viable cells, induced G2/M phase arrest and apoptosis based on sub‐G1 phase development. Furthermore, SFN increased ROS and Ca 2+ production and decreased the levels of ΔΨ m and activated caspase‐3, −8, and −9 activities in HL‐60 cells. SFN significantly upregulated the expression of BAX, Bid, Fas, Fas‐L, caspase‐8, Endo G, AIF, and cytochrome c, and inhibited the antiapoptotic proteins such as Bcl‐x and XIAP, that is associated with apoptosis. We also used cDNA microarray to confirm several gene expressions such as caspase −8, −3, −4, −6, and −7 that are affected by SFN. Those results indicated that SFN induced apoptosis in HL‐60 cells via Fas‐ and mitochondria‐dependent pathways. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 311–328, 2017.