Abstract
Aberrant activation of the Wnt/β-catenin signaling pathway is a common event in human tumor progression. Wnt signaling has also been implicated in maintaining a variety of adult and embryonic stem cells by imposing a restraint to differentiation. To understand the function and mechanism of Wnt/β-catenin signaling on the pathogenesis of teratocarcinoma, we used the mouse teratocarcinoma P19 cell line as a model in vitro. Gsk3β specific inhibitor (SB216763) was used to activate Wnt/β-catenin signaling. All trans-retinoic acid (RA) was used to induce P19 cell differentiation. At different culture times, gene expression was examined by immunofluorescence staining, quantitative real-time PCR, and Western-blotting; BrdU incorporation assays were performed to measure P19 cell proliferation. Small interference RNA technology was used to downregulate c-myc expression. The results showed that SB216763 induced the nuclear translocation of β-catenin, upregulated the expression of c-myc and pluripotency related genes, oct4, sox2 and nanog, and blocked cell differentiation induced by all trans-RA. The proliferation of P19 cells was significantly enhanced by SB216763, as well as c-myc overexpression. C-myc downregulation inhibited P19 cell proliferation caused by activation of Wnt/β-catenin signaling and induced P19 cell differentiation. In conclusion, activation of the Wnt/β-catenin pathway could promote the proliferation and inhibit the differentiation of mouse teratocarcinoma cells by upregulation of c-myc expression.
Copyright © 2012 Wiley Periodicals, Inc.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Blotting, Western
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Cell Differentiation / drug effects
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Cell Line, Tumor
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Cell Proliferation* / drug effects
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Embryonal Carcinoma Stem Cells / drug effects
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Embryonal Carcinoma Stem Cells / metabolism*
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Embryonal Carcinoma Stem Cells / pathology
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Fluorescent Antibody Technique
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Gene Expression Regulation, Developmental
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Gene Expression Regulation, Neoplastic
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Glycogen Synthase Kinase 3 / antagonists & inhibitors
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Glycogen Synthase Kinase 3 / metabolism
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Glycogen Synthase Kinase 3 beta
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Homeodomain Proteins / genetics
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Homeodomain Proteins / metabolism
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Indoles / pharmacology
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Maleimides / pharmacology
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Mice
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Nanog Homeobox Protein
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Octamer Transcription Factor-3 / genetics
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Octamer Transcription Factor-3 / metabolism
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Protein Kinase Inhibitors / pharmacology
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Proto-Oncogene Proteins c-myc / genetics
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Proto-Oncogene Proteins c-myc / metabolism*
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RNA Interference
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RNA, Messenger / metabolism
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Real-Time Polymerase Chain Reaction
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SOXB1 Transcription Factors / genetics
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SOXB1 Transcription Factors / metabolism
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Teratocarcinoma / genetics
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Teratocarcinoma / metabolism*
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Teratocarcinoma / pathology
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Time Factors
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Transfection
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Tretinoin / pharmacology
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Up-Regulation
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Wnt Signaling Pathway* / drug effects
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beta Catenin / metabolism*
Substances
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CTNNB1 protein, mouse
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Homeodomain Proteins
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Indoles
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Maleimides
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Myc protein, mouse
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Nanog Homeobox Protein
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Nanog protein, mouse
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Octamer Transcription Factor-3
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Pou5f1 protein, mouse
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Protein Kinase Inhibitors
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Proto-Oncogene Proteins c-myc
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RNA, Messenger
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SB 216763
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SOXB1 Transcription Factors
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Sox2 protein, mouse
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beta Catenin
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Tretinoin
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GSK3B protein, human
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Glycogen Synthase Kinase 3 beta
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Gsk3b protein, mouse
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Glycogen Synthase Kinase 3