OBJECTIVE: Prostate-specific antigen measurement, widely used for early detection of prostate cancer (CaP), suffers from low specificity. Additional tumor markers are needed for the early detection of clinically relevant CaP. Our objective was to perform a qualitative proteomic analysis of conditioned medium (CM) from the CaP cell line PC3(AR)(6). METHODS: We used a roller bottle culture system to culture the PC3(AR)(6) cell line in chemically defined serum-free medium for 14 days. By using strong anion-exchange chromatography, we fractionated the CM and trypsinized the fractions. The tryptic peptides were further fractionated by reversed-phase C-18 chromatography before being subjected to electrospray ionization tandem mass spectrometry. We used MASCOT software to search the mass spectra generated and organized identified proteins based on their genome ontology classification of cellular location. We used an immunoassay to measure a newly identified secreted protein, Mac-2BP, and kallikreins 5, 6, and 11 in serum samples from CaP patients and healthy men. RESULTS: We classified 262 proteins according to cellular location; the sample was found to contain a significant proportion of secreted (23%) and membrane (16%) proteins. In a proportion of cancer patients compared with healthy men, we determined by ELISA that serum concentrations of a novel candidate biomarker Mac-2BP were increased. CONCLUSIONS: These identified proteins, and possibly many others found in the CM, may have utility as novel CaP biomarkers.
OBJECTIVE:Prostate-specific antigen measurement, widely used for early detection of prostate cancer (CaP), suffers from low specificity. Additional tumor markers are needed for the early detection of clinically relevant CaP. Our objective was to perform a qualitative proteomic analysis of conditioned medium (CM) from the CaP cell line PC3(AR)(6). METHODS: We used a roller bottle culture system to culture the PC3(AR)(6) cell line in chemically defined serum-free medium for 14 days. By using strong anion-exchange chromatography, we fractionated the CM and trypsinized the fractions. The tryptic peptides were further fractionated by reversed-phase C-18 chromatography before being subjected to electrospray ionization tandem mass spectrometry. We used MASCOT software to search the mass spectra generated and organized identified proteins based on their genome ontology classification of cellular location. We used an immunoassay to measure a newly identified secreted protein, Mac-2BP, and kallikreins 5, 6, and 11 in serum samples from CaP patients and healthy men. RESULTS: We classified 262 proteins according to cellular location; the sample was found to contain a significant proportion of secreted (23%) and membrane (16%) proteins. In a proportion of cancerpatients compared with healthy men, we determined by ELISA that serum concentrations of a novel candidate biomarker Mac-2BP were increased. CONCLUSIONS: These identified proteins, and possibly many others found in the CM, may have utility as novel CaP biomarkers.
Authors: Eric W Klee; Olga P Bondar; Marcia K Goodmanson; Roy B Dyer; Sibel Erdogan; Eric J Bergstralh; H Robert Bergen; Thomas J Sebo; George G Klee Journal: Clin Chem Date: 2012-01-12 Impact factor: 8.327
Authors: Kerry L Inder; Yu Zi Zheng; Melissa J Davis; Hyeongsun Moon; Dorothy Loo; Hien Nguyen; Judith A Clements; Robert G Parton; Leonard J Foster; Michelle M Hill Journal: Mol Cell Proteomics Date: 2011-10-26 Impact factor: 5.911
Authors: Rajaventhan Srirajaskanthan; Martyn E Caplin; Mark G Waugh; Jennifer Watkins; Tim Meyer; J Justin Hsuan; Nicholas J Beaumont Journal: Mol Cell Proteomics Date: 2009-12-17 Impact factor: 5.911
Authors: Susanne Klein-Scory; Salwa Kübler; Hanna Diehl; Christina Eilert-Micus; Anke Reinacher-Schick; Kai Stühler; Bettina Warscheid; Helmut E Meyer; Wolff Schmiegel; Irmgard Schwarte-Waldhoff Journal: BMC Cancer Date: 2010-02-25 Impact factor: 4.430
Authors: John D Andersen; Kristin Lm Boylan; Ronald Jemmerson; Melissa A Geller; Benjamin Misemer; Katherine M Harrington; Starchild Weivoda; Bruce A Witthuhn; Peter Argenta; Rachel Isaksson Vogel; Amy Pn Skubitz Journal: J Ovarian Res Date: 2010-09-10 Impact factor: 4.234