Proteomic Analysis Of Differential Expression Proteins In Lung Adenocarcinoma Cell Line(A549) And Resistand Cell Line(A549/DDP)

Lung cancer is one of the most malignant tumor with high morbidity and mortality,as a threat to human health all over the world at present. The fatality of the cancer deaths accounted for about 17.6%,five-year survival rate of only 8.9%—15%with a gradual upward trend.China will become one of the countries which possess the highest incidence in the twenty-first century estimated by WHO.Adenocarcinoma as the most common type of Lung cancer,its treatment is a surgery, chemotherapy-based integrated multi-disciplinary treatment,including chemotherapy for the treatment of one of the important means,but the current indicators of chemotherapy patients who through chemotherapy has efficiency only about 30 percent more than,its five-year survival rate of less than 15%.The main reason for the failure of chemotherapy is the drug-resistance caused in the course of chemotherapy.The resistance mechanisms are the result of combined effect of many factors and genes. The previous studies of drug resistance of lung cancer tended to be concentrated in a single or a few related genes and their protein products on,which is difficult to reflect the changing picture of lung cancer DNA.Purpose:Directly Focus on the total protein,with the method of proteome with the discrepancy to find the drug-resistance related point of the protein of the Adenocarcinoma of lung.It might bring light to the process and the essence of the Adenocarcinoma of lung comprehensively and literally,and inaugurat the new path and clew and also the theory for the clinic pharmace,curative effect,and the targeting point,as well as for the prediction of clinical prognosis of lung cancer metastasis and recurrence and provide.Methodcultivate and collecte the samples of drug-resistant lung adenocarcinoma(A549/DDP) and non-drug-resistant(A549) cell lines, and separate the total protein patterns with the two-dimensional gel electrophoresis,PDQuest software applications on the two-dimensional gel electrophoresis map of the differences in the expression of more than 2 times the protein to carry out a comparative analysis using matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS) for differential expression analysis of proteins,to obtain MALDI-TOF-MS peptide mass fingerprint database search identified 20 differentially expressed protein non-redundant,then identify part of the discrepancy of the protein with the data of the bioinformatics.ResultObtain the sample and detect about 1000 points in each line after Coomassie brilliant blue,then pick up most distinctive 40 points for the in the MOLDI-TOF-MS mass spectra identification in the 100 differences by PDQuest software applications on the two-dimensional gel electrophoresis map of the differences in the expression of more than 2 times the protein to carry out a comparative analysis using matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS) for differential expression analysis of proteins,to obtain MALDI-TOF-MS peptide mass fingerprint database search identified 20 differentially expressed protein non-redundant.35 peptide mass fingerprinting are reveal in the mass spectra identification,22 of them are approved in the database,and 2 are duplicated;5 are null.There are 20 points identified in the initial tests by Mascot software in the SwiSS-PROT databates,Such as Heat shock protein beta27, Annexin A2,Histone H4,Triosephosphate isomerase,fatty acid-binding proteins(FABP),Peroxinredoxin-4,Elongation factor-1Conclusion(1) Separate the total protein pattern and get the map with the two-dimensional gel electrophoresis of the drug-resistant lung adenocarcinoma(A549/DDP) and non-drug-resistant(A549) cell fitly.(2) the expression of the protein of drug-resistant lung adenocarcinoma(A549/DDP) and non-drug-resistant(A549) cell distincts conspicuously.By the way of discrepancy of the proteome,it successfully identified the targeting protein,and is useful for the the foundation of the database of the adenocarcinoma of lung.(3) the protein,such as Heat shock protein beta27,Annexin A2, Histone H4,Triosephosphate isomerase,fatty acid-binding proteins (FABP),Peroxinredoxin-4,Elongation factor-1,probablely are the special protein related to the drug-resistance,which might be for the further study of the func-tions of the protein.