| Objective: The study aims to research the proteomic differences among the high and the low differentiation cancer of larynx tissues and the normal mucous epithelium by using two--dimensional gel electrophoresis(2-DE) and mass spectrometry.Methods: In sample preparation,the fresh carcinoma tissue was cut of from the specimen immediately after the specimen was resected from the body. The tissues were classified into three groups according to the postoperative pathological diagnosis:high differentiation carcinoma ,low differentiation carcinoma and normal epithelium.In the first part of our experiment,protein separating Was performed using two—dimensional gel electrophoresis(2-DE) with 17cm linear pH 3-10 immobilized pH gradient(IPG)strips.After Coomassie brilliant blue—staining, the images of the gels were acquired by the scanner and then analyzed to find the differentially expressed protein—spots in each group.In the second part of the experiment,the differentially expressed protein—spots Was incised from the gels and digested by trypsin.We acquired the peptide mass fingerprintings(PMF) by matrix assisted laser desorption /ionization time 一of—flight mass spectrometry(MALDI_T0F—MS)and identified the proteins by data searching in the MS-Fit database.Results: eleven proteins were preliminarily identified. We detected Cofilin-1,Cyclophilin A expressed exclusively in both the two cancer groups .In high differentiation group,S100 calcium-binding protein A11 and RAF proto-oncogene serine/threonine-protein kinase expressed exclusively and Nuclear body protein SP140 and Aryl hydrocarbon receptor nuclear translocator 2 increased .In low differentiation group,glucose-regulated protein (GRP94) ,Zinc finger protein 624, Glutathione S-transferase P , ADP/ATP translocase 3 and Centromere protein S (CENP-S) increased.Conclusion: Some differentially expressed proteins existed among high and the low differentiation carcinoma of larynx as well as the normal laryngeal mucosa. |
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