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Comparison of intracellular signalling by insulin and the hypermitogenic AspB10 analogue in MCF-7 breast adenocarcinoma cells.
MedLine Citation:
PMID:  20936651     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
We compared mitogenicity and intracellular signalling by human insulin and the AspB10 (X-10) human insulin analogue in MCF-7 human mammary adenocarcinoma cells. By flow analysis of phosphorylated histone H3 or cell cycle distributions, insulin and X-10 were mitogenic at physiologically relevant concentrations (2 nm to 74 pm range), with X-10 being approximately 3-fold more mitogenic than insulin. By western blotting with phospho-specific antibodies, insulin induced phosphorylation of IRS-1, Akt, p70S6K, S6 ribosomal protein, 4E-BP1, FoxO3a, FoxO1, p44/42 MAPK and the EGFR. Blocking with wortmannin, rapamycin and U0126 showed that these signalling events conformed to the canonical PI3K pathway. IRS-1 (Ser302) phosphorylation was abolished by wortmannin and rapamycin, suggesting a feedback from the PI3K pathway on insulin signalling. Compared with equimolar insulin, X-10 caused up to 2-fold higher phosphorylation of all proteins examined in this study. The phosphorylation sites that responded most strongly to insulin were not generally the same as those responding most strongly to X-10. In the PI3K pathway, the most X-10-sensitive protein localized to the translation-regulating arm (p70S6K), with FoxO3a and FoxO1 transcription factors showing a more comparable response to insulin and X-10. Using flow analysis, we confirmed the correlation between insulin-triggered translational activation in G0/G1 (S6 phosphorylation) and S-phase entry by MCF-7 cells. In summary, our findings implicate asymmetrical PI3K pathway activation and specifically stimulation of protein translation in the hypermitogenic effect of insulin analogues such as X-10. It remains to be shown whether these findings are relevant to other human mammary cancer cell types. Copyright © 2010 John Wiley & Sons, Ltd.
Authors:
Martin B Oleksiewicz; Christine Bonnesen; Anne Charlotte Hegelund; Anders Lundby; Gitte-Mai Nelander Holm; Marianne B Jensen; Jonas S Krabbe
Publication Detail:
Type:  Journal Article     Date:  2010-10-08
Journal Detail:
Title:  Journal of applied toxicology : JAT     Volume:  31     ISSN:  1099-1263     ISO Abbreviation:  J Appl Toxicol     Publication Date:  2011 May 
Date Detail:
Created Date:  2011-05-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8109495     Medline TA:  J Appl Toxicol     Country:  England    
Other Details:
Languages:  eng     Pagination:  329-41     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 John Wiley & Sons, Ltd.
Affiliation:
Molecular Toxicology, Novo Nordisk A/S, Novo Nordisk Park, DK-2760, Maalov, Denmark. mboleks@gmail.com.
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