Quantitative proteome analysis of ovarian cancer tissues using a iTRAQ approach

Abstract

Quantitative proteomics can be used as a screening tool for identification of differentially expressed proteins as potential biomarkers for cancers. Here, we comparatively analyzed the proteome profiles of ovarian cancer tissues and normal ovarian epithelial tissues. Using the high-throughput proteomic technology of isobaric tags for relative and absolute quantitation (iTRAQ)-coupled with two-dimensional-liquid chromatography-tandem mass spectrometry, 1,259 unique proteins were identified. Of those, 205 were potentially differentially expressed between ovarian cancer and normal ovarian tissues. Several of the potentially differentially expressed proteins were validated by Western blotting and real-time quantitative RT-PCR analyses. Furthermore, up-regulation of KRT8, PPA1, IDH2, and S100A11 were validated in ovarian tissue microarrays by immunohistochemistry. Silencing of S100A11 expression suppressed the migration and invasion properties of ovarian cancer cells in vitro. Our study represents the successful application of iTRAQ technology to an investigation of ovarian cancer. Many of the potentially differentially expressed proteins identified had not been linked to ovarian cancer before, and provide valuable novel insights into the underlying mechanisms of carcinogenesis in human ovarian cancer.

Proteomics Analysis of Ovarian Cancer Cell Lines and Tissues Reveals Drug Resistance-associated Proteins

“…Out of these nine proteins, expression of alpha-1-antichymotrypsin (AACT), fatty acid-binding protein, epidermal (FABP5), galectin-7 (LEG7), haemoglobin subunit beta (HBB), Ig gamma-1 chain C region (IGHG1) and serpin B3 (SPB3) were reduced, whilst EF1A1 and G3P, were increased in expression. In support of our findings, Wang and co-workers reported EF1A1 and G3P to be up-regulated in a quantitative proteome analysis of ovarian cancer tissues, while HBB was down-regulated (42).…”

“…Some of them [60 kDa heat-shock protein (CH60), ENOA, cathepsin D (CATD), EFTU, G3P and GRP75] were confirmed by a quantitative proteomic analysis using an iTRAQ approach (42). Interestingly, APOA1 and PRDX2 have also been found lobe up-regulated in ovarian cyst fluid of patients diagnosed with advanced serous ovarian adenocarcinoma (43).…”

Tissue Proteomics for the Next Decade? Towards a Molecular Dimension in Histology

“…LC-MS based quantitative methods such as label free quantification (Higgs et al, 2005;Neilson et al, 2011;Wang et al, 2006), spectral counting quantification (Neilson et al, 2011), intensity based quantification (Bondarenko et al, 2002;Christin et al, 2011), and also labeling techniques were developed for off-gel analyses. Labeling methods such as isobaric tag for relative and absolute quantification (iTRAQ) (Aggarwal et al, 2006;Chen et al, 2010;DeSouza et al, 2005;Garbis et al, 2008;Muraoka et al, 2012;Wang et al, 2012), and the recently developed isotope coded protein labeling coupled to immunoprecipitation (ICPL-IP) (Vogt et al, 2013), both relying on the labeling of the primary amino groups and lysine residues, were developed. Isotope coded affinity tag (ICAT) (DeSouza et al, 2005), that labels cysteine residues was also applied for tissue proteomics.…”

Identification and validation of differentially expressed proteins in epithelial ovarian cancers using quantitative proteomics

“…Wang et al . [6] found KRT8 protein, inorganic pyrophosphatase (PPA1), isocitrate dehydrogenase (IDH2), and protein S100-A11 were up-regulated in ovarian cancer tissue using quantitative proteome analysis with an iTRAQ approach. Also, Calcium-Activated Chloride Channel Regulator 1 (CLCA1) was revealed to be overexpressed in ovarian cancer cell [7].…”