title: 2-Methoxyestradiol induces endoreduplication through the induction of mitochondrial oxidative stress and the activation of MAPK signaling pathways.
abstract: 2-Methoxyestradiol (2ME2) is a normal physiological metabolite of 17beta-estradiol with anti-proliferative and anti-angiogenic activities. The purpose of this study is to elucidate the mechanism whereby 2ME2 induces endoreduplication of the well-differentiated nasopharyngeal carcinoma (NPC) cells. We report here that 2ME2 induces G2/M phase cell cycle arrest followed by endoreduplication of the well-differentiated HK-1 cells. The increase in chromosome number was confirmed by cytogenetic study. Analysis of stress signaling pathways revealed the phosphorylation activation of ERK, JNK and p38 MAPKs at various times after 2ME2 treatment. Pre-treatment of 2ME2-treated HK-1 cells with JNK inhibitor (SP600125), ERK inhibitor (PD98059) and p38 MAPK inhibitor (SB203580) resulted in the reduction of endoreduplicating cells. Furthermore, the increase in the phosphorylation of JNK was accompanied by an increase in the reactive oxygen species. In addition, endoreduplication was observed in cells after treatment with superoxide donor, 2,3-dimethoxy-1,4-naphoquinone (DMNQ). Confocal microscopic analysis also revealed the increase in mitochondrial superoxide anion in 2ME2-treated HK-1 cells. Pre-treatment of HK-1 cells with superoxide dismutase mimetic 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) or overexpressing the mitochondrial enzyme MnSOD resulted in the reduction of phosphorylation of JNK and the formation of endoreduplicating cells. Furthermore, the tubulin filaments in cytoplasm remain intact in 2ME2-treated HK-1 cells after pre-treatment of TEMPO. Our results suggest that 2ME2 induces endoreduplication through the induction of oxidative stress and the activation of MAPK signal pathways. The biological significance of drug-induced endoreduplication will also be discussed.
authors.affiliation: Department of Biology, Hong Kong Baptist University, 224, Waterloo Road, Hong Kong.
authors.forename: C M
authors.initials: CM
authors.lastname: Ting
authors.forename: Y M
authors.initials: YM
authors.lastname: Lee
authors.forename: C K C
authors.initials: CK
authors.lastname: Wong
authors.forename: A S
authors.initials: AS
authors.lastname: Wong
authors.forename: H L
authors.initials: HL
authors.lastname: Lung
authors.forename: M L
authors.initials: ML
authors.lastname: Lung
authors.forename: K W
authors.initials: KW
authors.lastname: Lo
authors.forename: R N S
authors.initials: RN
authors.lastname: Wong
authors.forename: N K
authors.initials: NK
authors.lastname: Mak
mesh_terms: D000077584:2-Methoxyestradiol; D002460:Cell Line; D049109:Cell Proliferation; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D050257:Tubulin Modulators / Q000494:pharmacology*
chemical_list: D050257:Tubulin Modulators; D004958:Estradiol; D000077584:2-Methoxyestradiol