title: A panel of regulated proteins in serum from patients with cervical intraepithelial neoplasia and cervical cancer.
abstract: We developed a discovery-validation mass-spectrometry-based pipeline to identify a set of proteins that are regulated in serum of patients with cervical intraepithelial neoplasia (CIN) and squamous cell cervical cancer using iTRAQ, label-free shotgun, and targeted mass-spectrometric quantification. In the discovery stage we used a "pooling" strategy for the comparative analysis of immunodepleted serum and revealed 15 up- and 26 down-regulated proteins in patients with early- (CES) and late-stage (CLS) cervical cancer. The analysis of nondepleted serum samples from patients with CIN, CES, an CLS and healthy controls showed significant changes in abundance of alpha-1-acid glycoprotein 1, alpha-1-antitrypsin, serotransferrin, haptoglobin, alpha-2-HS-glycoprotein, and vitamin D-binding protein. We validated our findings using a fast UHPLC/MRM method in an independent set of serum samples from patients with cervical cancer or CIN and healthy controls as well as serum samples from patients with ovarian cancer (more than 400 samples in total). The panel of six proteins showed 67% sensitivity and 88% specificity for discrimination of patients with CIN from healthy controls, a stage of the disease where current protein-based biomarkers, for example, squamous cell carcinoma antigen (SCCA), fail to show any discrimination. Additionally, combining the six-protein panel with SCCA improves the discrimination of patients with CES and CLS from healthy controls.
authors.affiliation: Department of Analytical Biochemistry, University of Groningen , Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands.
authors.forename: Alexander P
authors.initials: AP
authors.lastname: Boichenko
authors.forename: Natalia
authors.initials: N
authors.lastname: Govorukhina
authors.forename: Harry G
authors.initials: HG
authors.lastname: Klip
authors.forename: A G J
authors.initials: AG
authors.lastname: van der Zee
authors.forename: Coşkun
authors.initials: C
authors.lastname: Güzel
authors.forename: Theo M
authors.initials: TM
authors.lastname: Luider
authors.forename: Rainer
authors.initials: R
authors.lastname: Bischoff
mesh_terms: D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D014408:Biomarkers, Tumor / Q000097:blood; D001798:Blood Proteins / Q000032:analysis* / Q000378:metabolism; D002294:Carcinoma, Squamous Cell / Q000097:blood / Q000378:metabolism / Q000473:pathology; D016022:Case-Control Studies; D005260:Female; D006801:Humans; D007553:Isotope Labeling; D008875:Middle Aged; D008969:Molecular Sequence Data; D040901:Proteomics / Q000379:methods; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D002583:Uterine Cervical Neoplasms / Q000097:blood* / Q000378:metabolism / Q000473:pathology; D002578:Uterine Cervical Dysplasia / Q000097:blood* / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D001798:Blood Proteins
keywords: LC−MS/MS; alpha-1-acid-glycoprotein; alpha-1-antitrypsin; alpha-2-HS-glycoprotein; cervical cancer; cervical intraepithelial neoplasia; haptoglobin; protein panel; serotransferrin; vitamin D-binding protein