Quantification of modified nucleosides in in vitro and in vivo samples of pancreatic Cancer by CE-UV and LC-MS: validation of bioanalytical methods

• Grant number: 13/13084-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: October 01, 2013
• End date: September 30, 2014
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
• Principal Investigator: Ana Valéria Colnaghi Simionato
• Grantee: Mariana de Oliveira Silva
• Host Institution: Instituto de Química (IQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Cancer is a malignant neoplasia that lead to the death of 7.6 million people in 2008 around the world. Its incidence has been growing and the predictions are alarming (13.1 million deaths in 2030). More than half of all cancer base pathologies can be cured when the early diagnosis is achieved, including the pancreatic, through surgery. This base pathology presents a high mortality index, because it is aggressive and do not show easily identifiable symptoms before an advanced state, usually metastasis. The lack of a noninvasive method for diagnosis in the beginning of the disease demonstrates how important the investigation of pancreatic tumor biomarkers is. The scientific literature shows that the concentration of modified nucleosides in biological fluids of sick individuals is bigger than in healthy ones, so they are potential tumor biomarkers. In this project electrophoretic methods and liquid chromatography coupled to mass spectrometry in tandem (LC-MS/MS) will be used to quantify modified nucleosides in serum and urine from patients with pancreatic cancer and healthy subjects. The supernatant from the in vitro samples obtained from the neoplastic cellular lines of pancreatic adenocarcinoma (Panc-1) and primary cellular line of three years old children fibroblast (3Y-P1), considering the last one as healthy cells, will be analyzed by the developed method and validate for comparative ends with the in vivo samples.The optimization of the chromatographic method will be performed, since the electrophoretic one has already been developed in our research group, and the validation of both will be carried out as well. Therefore it will be possible to analyze the presence of certain nucleosides as potential tumor biomarkers of pancreatic cancer, aiding the diagnosis of this base pathology through the corroboration with exams routinely done in clinical analyses laboratory, where immunoassays are performed for quantification of the carbohydrate antigen (CA 19-9), leading to a more reliable and earlier diagnostic, which could raise the survival rate of the patient.