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Different Changes in Protein and Phosphoprotein Levels Result from Serum Starvation of High-Grade Glioma and Adenocarcinoma Cell Lines

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Departments of Neuro-Oncology, Bioinformatics and Computational Biology, and Blood and Marrow Transplantation, Section of Molecular Hematology, The University of Texas M. D. Anderson Cancer Center, Houston, Texas 77230
* Author to whom correspondence should be addressed; Victor A. Levin, Department of Neuro-Oncology, Unit 431, The University of Texas M. D. Anderson Cancer Center, P.O. Box 301402, Houston, TX 77230-1402 USA. Tel: +1 713-792-8297. Fax: +1 713-794-4999. E-mail: [email protected]
†Department of Neuro-Oncology.
‡Department of Bioinformatics and Computational Biology.
§Department of Blood and Marrow Transplantation.
Cite this: J. Proteome Res. 2010, 9, 1, 179–191
Publication Date (Web):November 8, 2009
Copyright © 2009 American Chemical Society

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    Tumor cells undergoing serum starvation in vitro partially mimic metabolically stressed cells trying to adjust to a changed environment in vivo by inducing signal transduction and gene expression so that the tumor continues to grow. Our hypothesis is that the changes in protein and phosphoprotein levels after serum starvation may reflect the adapted phenotype of the tumor, which could be targeted for therapy. We used reverse-phase protein microarrays to interrogate five high-grade glioma cell lines and seven adenocarcinoma cell lines for differences in the level of 81 proteins and 25 phosphoproteins. All cell lines were studied in the well-fed condition of growth with 10% FBS and the starved condition of 0.5% FBS. Protein expression levels were normalized to β-actin and trichotomized as increased (+1, upper 75th quartile), decreased (−1, lowest 25th quartile), or unchanged (0, others) to focus on the patterns of the biggest (and hopefully most robust) changes in protein and phosphoprotein levels. We examined these trichotomized values to better understand Starved-Fed differences among the cell lines and thereby gain better/clearer insight into the effects of serum starvation on potential cellular responses. In general, the expression of proteins and phosphoproteins 24 h after FBS starvation increased more often in glioma lines than in adenocarcinoma lines, which appeared to have fewer increased protein scores and more decreased scores. Many of the proteins increased in gliomas were downstream targets of the PTEN-PI-3 kinase-AKT, EGFR-MAPK-Stat, and transcription activator-polyamine signaling pathways. In adenocarcinomas, the expression of proteins and phosphoproteins generally increased in apoptosis pathways, while there were minor fluctuations in the other pathways above. Contrawise, gliomas become resistant to apoptosis after 24 h of serum starvation and upregulate transcription activators and polyamines more so than adenocarciomas.

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