Cargando…

The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations

BACKGROUND: The role of Notch signaling in HPV-mediated transformation has been a long standing debate, as both tumor suppressive and oncogenic properties have been described. We examined whether the dual findings in literature may be explained by gene dosage effects and determined the relation with...

Descripción completa

Detalles Bibliográficos
Autores principales: Henken, Florianne E., De-Castro Arce, Johanna, Rösl, Frank, Bosch, Leontien, Meijer, Chris J. L. M., Snijders, Peter J. F., Steenbergen, Renske D. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306567/
https://www.ncbi.nlm.nih.gov/pubmed/22271085
http://dx.doi.org/10.1007/s13402-011-0062-4
_version_ 1782227229798825984
author Henken, Florianne E.
De-Castro Arce, Johanna
Rösl, Frank
Bosch, Leontien
Meijer, Chris J. L. M.
Snijders, Peter J. F.
Steenbergen, Renske D. M.
author_facet Henken, Florianne E.
De-Castro Arce, Johanna
Rösl, Frank
Bosch, Leontien
Meijer, Chris J. L. M.
Snijders, Peter J. F.
Steenbergen, Renske D. M.
author_sort Henken, Florianne E.
collection PubMed
description BACKGROUND: The role of Notch signaling in HPV-mediated transformation has been a long standing debate, as both tumor suppressive and oncogenic properties have been described. We examined whether the dual findings in literature may be explained by gene dosage effects and determined the relation with AP-1, a downstream target of Notch. METHODS: SiHa cervical cancer cells were transfected with two doses of intracellular active Notch. Non-tumorigenic HPV16-immortalized keratinocytes (FK16A) were transfected with Fra1 specific siRNAs and non-targeting controls. Transfectants were analysed for Notch, Hes, cJun, cFos and Fra1 mRNA expression, Notch pathway activation using luciferase assays, cell viability using MTT assays, anchorage independent growth, AP-1 activity and/or AP-1 complex composition using EMSA. RESULTS: In SiHa cells two activation states of Notch signaling pathway were obtained. Moderate Notch activation contributed to increased viability and anchorage independent growth, whereas high level Notch activation decreased anchorage independent growth. The shift in phenotypical outcome was correlated to altered AP-1 activity and complex composition. Moderate Notch expression led to an increased AP-1 transcriptional activity and DNA binding activity, but did not affect complex composition. High levels of Notch additionally led to a change in AP-1 complex composition, from cJun/cFos to cJun/Fra1 dimers, which is exemplary for non-tumorigenic HPV-immortalized cell lines. Conversely, silencing of Fra1 in non-tumorigenic HPV16-immortalized keratinocytes, leading to an enrichment of cJun/cFos dimers, was accompanied with increased colony formation. CONCLUSION: The functional role of Notch in HPV-mediated transformation is dosage dependent and correlated to a change in AP-1.
format Online
Article
Text
id pubmed-3306567
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher Springer Netherlands
record_format MEDLINE/PubMed
spelling pubmed-33065672012-03-22 The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations Henken, Florianne E. De-Castro Arce, Johanna Rösl, Frank Bosch, Leontien Meijer, Chris J. L. M. Snijders, Peter J. F. Steenbergen, Renske D. M. Cell Oncol (Dordr) Original Paper BACKGROUND: The role of Notch signaling in HPV-mediated transformation has been a long standing debate, as both tumor suppressive and oncogenic properties have been described. We examined whether the dual findings in literature may be explained by gene dosage effects and determined the relation with AP-1, a downstream target of Notch. METHODS: SiHa cervical cancer cells were transfected with two doses of intracellular active Notch. Non-tumorigenic HPV16-immortalized keratinocytes (FK16A) were transfected with Fra1 specific siRNAs and non-targeting controls. Transfectants were analysed for Notch, Hes, cJun, cFos and Fra1 mRNA expression, Notch pathway activation using luciferase assays, cell viability using MTT assays, anchorage independent growth, AP-1 activity and/or AP-1 complex composition using EMSA. RESULTS: In SiHa cells two activation states of Notch signaling pathway were obtained. Moderate Notch activation contributed to increased viability and anchorage independent growth, whereas high level Notch activation decreased anchorage independent growth. The shift in phenotypical outcome was correlated to altered AP-1 activity and complex composition. Moderate Notch expression led to an increased AP-1 transcriptional activity and DNA binding activity, but did not affect complex composition. High levels of Notch additionally led to a change in AP-1 complex composition, from cJun/cFos to cJun/Fra1 dimers, which is exemplary for non-tumorigenic HPV-immortalized cell lines. Conversely, silencing of Fra1 in non-tumorigenic HPV16-immortalized keratinocytes, leading to an enrichment of cJun/cFos dimers, was accompanied with increased colony formation. CONCLUSION: The functional role of Notch in HPV-mediated transformation is dosage dependent and correlated to a change in AP-1. Springer Netherlands 2012-01-24 2012 /pmc/articles/PMC3306567/ /pubmed/22271085 http://dx.doi.org/10.1007/s13402-011-0062-4 Text en © The Author(s) 2012 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Original Paper
Henken, Florianne E.
De-Castro Arce, Johanna
Rösl, Frank
Bosch, Leontien
Meijer, Chris J. L. M.
Snijders, Peter J. F.
Steenbergen, Renske D. M.
The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations
title The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations
title_full The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations
title_fullStr The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations
title_full_unstemmed The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations
title_short The functional role of Notch signaling in HPV-mediated transformation is dose-dependent and linked to AP-1 alterations
title_sort functional role of notch signaling in hpv-mediated transformation is dose-dependent and linked to ap-1 alterations
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3306567/
https://www.ncbi.nlm.nih.gov/pubmed/22271085
http://dx.doi.org/10.1007/s13402-011-0062-4
work_keys_str_mv AT henkenfloriannee thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT decastroarcejohanna thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT roslfrank thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT boschleontien thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT meijerchrisjlm thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT snijderspeterjf thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT steenbergenrenskedm thefunctionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT henkenfloriannee functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT decastroarcejohanna functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT roslfrank functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT boschleontien functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT meijerchrisjlm functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT snijderspeterjf functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations
AT steenbergenrenskedm functionalroleofnotchsignalinginhpvmediatedtransformationisdosedependentandlinkedtoap1alterations