Proteomic analysis of temporally stimulated ovarian cancer cells for biomarker discovery

Research output: Contribution to journalArticle

Abstract

While ovarian cancer remains the most lethal gynecological malignancy in the United States, there are no biomarkers available that are able to predict therapeutic responses to ovarian malignancies. One major hurdle in the identification of useful biomarkers has been the ability to obtain enough ovarian cancer cells from primary tissues diagnosed in the early stages of serous carcinomas, the most deadly subtype of ovarian tumor. In order to detect ovarian cancer in a state of hyperproliferation, we analyzed the implications of molecular signaling cascades in the ovarian cancer cell line OVCAR3 in a temporal manner, using a mass-spectrometry-based proteomics approach. OVCAR3 cells were treated with EGF1, and the time course of cell progression was monitored based on Akt phosphorylation and growth dynamics. EGF-stimulated Akt phosphorylation was detected at 12 h posttreatment, but an effect on proliferation was not observed until 48 h post-exposure. Growth-stimulated cellular lysates were analyzed for protein profiles between treatment groups and across time points using iTRAQ labeling and mass spectrometry. The protein response to EGF treatment was identified via iTRAQ analysis in EGF-stimulated lysates relative to vehicle-treated specimens across the treatment time course. Validation studies were performed on one of the differentially regulated proteins, lysosomal-associated membrane protein 1 (LAMP-1), in human tissue lysates and ovarian tumor tissue sections. Further, tissue microarray analysis was performed to demarcate LAMP-1 expression across different stages of epithelial ovarian cancers. These data support the use of this approach for the efficient identification of tissuebased markers in tumor development related to specific signaling pathways. LAMP-1 is a promising biomarker for studies of the progression of EGF-stimulated ovarian cancers and might be useful in predicting treatment responses involving tyrosine kinase inhibitors or EGF receptor monoclonal antibodies.

Original languageEnglish (US)
Pages (from-to)356-368
Number of pages13
JournalMolecular and Cellular Proteomics
Volume12
Issue number2
DOIs
StatePublished - Feb 2013

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Lysosomal-Associated Membrane Protein 1
Biomarkers
Tumor Biomarkers
Epidermal Growth Factor
Proteomics
Ovarian Neoplasms
Cells
Tissue
Tumors
Phosphorylation
Mass spectrometry
Mass Spectrometry
Neoplasms
Tissue Array Analysis
Microarrays
Epidermal Growth Factor Receptor
Proteins
Protein-Tyrosine Kinases
Labeling
Validation Studies

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Cite this

Proteomic analysis of temporally stimulated ovarian cancer cells for biomarker discovery. / Marzinke, Mark A; Choi, Caitlin H.; Chen, Li; Shih, Ie Ming; Chan, Daniel Wan-Yui; Zhang, Hui.

In: Molecular and Cellular Proteomics, Vol. 12, No. 2, 02.2013, p. 356-368.

Research output: Contribution to journalArticle

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