BACKGROUND: Early diagnosis and treatment of gastric cancer patients is essential for improving prognosis. However, no available serum-based test provides sufficient sensitivity or specificity for widespread use. Therefore, in this study we aimed to identify cancer biomarkers in human sera using 2-dimensional difference gel electrophoresis (2D-DIGE), and to characterize protein biomarkers with tandem mass spectrometry. METHODS: We compared the serum proteomic profiles of 20 gastric cancer patients and 10 healthy volunteers. Serum samples were first chromatographed using an immunoaffinity high-performance liquid chromatography (HPLC) column to selectively remove albumin, immunoglobulins, transferrin, haptoglobin, and antitrypsin. Differential protein analysis was then performed using DIGE. Significantly increased and decreased protein spot features were excised, trypsin digested, and analyzed by tandem matrix-assisted laser desorption/ionization (MALDI) time of flight (TOF)/TOF and a linear trap quadrupole (LTQ) mass spectrometer. RESULTS: Seventeen protein spot features were significantly increased and 7 were significantly decreased in cancer serum samples compared to healthy controls. We identified 7 unique proteins that were upregulated, including plasminogen, apolipoprotein A-IV, Kininogen-1, complex-forming glycoprotein HC, complement component C4A, apolipoprotein J, and clusterin, and 5 that were decreased. CONCLUSIONS: These results suggest that the combination of multi-dimensional HPLC and 2D-DIGE provides a valuable tool for serum proteomics in gastric cancer.
BACKGROUND: Early diagnosis and treatment of gastric cancerpatients is essential for improving prognosis. However, no available serum-based test provides sufficient sensitivity or specificity for widespread use. Therefore, in this study we aimed to identify cancer biomarkers in human sera using 2-dimensional difference gel electrophoresis (2D-DIGE), and to characterize protein biomarkers with tandem mass spectrometry. METHODS: We compared the serum proteomic profiles of 20 gastric cancerpatients and 10 healthy volunteers. Serum samples were first chromatographed using an immunoaffinity high-performance liquid chromatography (HPLC) column to selectively remove albumin, immunoglobulins, transferrin, haptoglobin, and antitrypsin. Differential protein analysis was then performed using DIGE. Significantly increased and decreased protein spot features were excised, trypsin digested, and analyzed by tandem matrix-assisted laser desorption/ionization (MALDI) time of flight (TOF)/TOF and a linear trap quadrupole (LTQ) mass spectrometer. RESULTS: Seventeen protein spot features were significantly increased and 7 were significantly decreased in cancer serum samples compared to healthy controls. We identified 7 unique proteins that were upregulated, including plasminogen, apolipoprotein A-IV, Kininogen-1, complex-forming glycoprotein HC, complement component C4A, apolipoprotein J, and clusterin, and 5 that were decreased. CONCLUSIONS: These results suggest that the combination of multi-dimensional HPLC and 2D-DIGE provides a valuable tool for serum proteomics in gastric cancer.
Authors: Yashwanth Subbannayya; Sartaj Ahmad Mir; Santosh Renuse; Srikanth S Manda; Sneha M Pinto; Vinuth N Puttamallesh; Hitendra Singh Solanki; H C Manju; Nazia Syed; Rakesh Sharma; Rita Christopher; M Vijayakumar; K V Veerendra Kumar; T S Keshava Prasad; Girija Ramaswamy; Rekha V Kumar; Aditi Chatterjee; Akhilesh Pandey; Harsha Gowda Journal: J Proteomics Date: 2015-05-05 Impact factor: 4.044
Authors: Pål Vange; Torunn Bruland; Berit Doseth; Reidar Fossmark; Mirta M L Sousa; Vidar Beisvag; Øystein Sørdal; Gunnar Qvigstad; Helge L Waldum; Arne K Sandvik; Ingunn Bakke Journal: PLoS One Date: 2017-09-13 Impact factor: 3.240