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The Extraction and Application of Endogenous Serum Phosphopeptides from Lung Cancer Patients

Author: LiJuan
Tutor: ZouLiJuan
School: Dalian Medical University
Course: Oncology
Keywords: Proteomics Serum Phosphorylation of endogenous peptides Tumor markers
CLC: R734.2
Type: Master's thesis
Year: 2011
Downloads: 20
Quote: 0
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Abstract


Background: The tumor is due to a group of proteins changes cause the entire signal path eventually occur abnormal and a proteomic disease. Looking tumor biomarkers for early diagnosis, pathological type, the development of new cancer drug targets, drug intervention is of great value, to become one of the main direction of proteomics of lung cancer in half a century increased morbidity and mortality fast, hazards to human health and life of the largest cancer, cancer causes of death in our country has leapt first place. Currently screening for early lung cancer is the primary means of imaging and sputum cytology, which there is a certain misdiagnosed, and pathological examination in clinical diagnosis as the gold standard but there is a big disadvantage for trauma patients. Therefore, the majority of patients with lung cancer can lead to early detection, early treatment, the patient's quality of life and survival rate of survival is affected. With the development of molecular biology, tumor markers has become a common means of diagnosis of malignant tumors of cancer screening has important clinical value. So, how to find lung cancer tumor markers has become proteomics research hotspot. Protein phosphorylation of proteins as one of the most extensive post-translational modification (Post Translational Modifi-cations, PTMs), and regulates the life activities in almost all the processes, including cell proliferation, cell differentiation, muscle contraction, nerve activity, metabolism, etc. . Role of abnormal phosphorylation of the root causes of human disease, one of the most significant disease is cancer. Therefore, the use of mass spectrometry for high sensitivity and rapid detection of phosphorylated proteins, according to the change of phosphorylated proteins to predict, judgment, diagnosis of disease, can greatly improve its accuracy, has important biological significance. Objective: In this experiment, serum acetone precipitation, uses a membrane to purify the supernatant after precipitation, and then use a metal oxide (Ti02) as enrichment materials, through a series of pre-treatment serum concentration United nano UPLC / Q-TOF proteomic analysis method to establish a serum endogenous phosphorylated peptides. By examining multiple sets of lung cancer patients and normal serum endogenous phosphorylated peptides difference proves that this method makes sense, for the identification of tumor markers in lung cancer provides a theoretical basis. Methods: As a standard sample, to establish a membrane filter, metal oxides enrichment and nanoliter ultrahigh-pressure liquid chromatography - mass spectrometry (nano UPLC-MS) combined with detection of phosphorylated peptides. Another 10 cases of lung cancer and 10 cases of normal serum aliquots, acetone precipitation and application of the above method established endogenous serum phosphorylated peptides. Results: 1. After exploration experimental conditions, the choice of filter membrane 10K, 50% CH3OH/0.5FA solvent to dissolve, 0.5μLα-adding 5μL serum casein as an experimental detection limit, established a proteomics phosphorylated peptide testing new methods. 2, with the establishment of serum proteomic methods to detect: 1. Sera from patients with lung cancer was detected in 14 of endogenous phosphorylated peptide was detected in 10 normal human serum endogenous phosphorylated peptides. Conclusion: Based on the lung cancer patients and normal serum endogenous phosphorylated peptide detection, identification, verification of successful established proteomics methods usability and value: a. Established a proteomics of complex samples treatment and detection theory methods. 2 Successful serum from patients with lung cancer was detected in 14 of endogenous phosphorylated peptides detected in normal human serum from 10 endogenous phosphorylated peptides. 3 of lung cancer patients and normal serum detected endogenous phosphorylated peptides verified by comparing the differences between the two, for the final identification of tumor markers in lung cancer foundation.

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CLC: > Medicine, health > Oncology > Respiratory system tumors > Lung tumors
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