Screening NPC Proteomics Technology Transfer-related Protein

BACKGROUND AND OBJECTIVESNasopharyngeal Carcinoma(NPC)is a human malignant tumor derived from nasopharyngeal epithelial cells, which is one of the ten highest incidences in our country and is the higest incidences of head and neck tumors. NPC is a secrete tumor and sometimes have lymph node metastasis or other organs metastasis before be found. So it is very important for therapy and prognosis of NPC patients to have early stage diagnose. But we have no early stage diagnose method of NPC until now. Therefore, it is very important to study NPC associated serum proteins by comparative proteome analysis. Proteomics technology have the features of high-flux and high sensitiveness, which can expresse different protein by real-time and by stage. This study provide the theory evidences for explaining metastasis mechanism of NPC and searching marker to predict NPC metastasis.METHODEwe culture NPC cells CNE1(well-differentiated squamous cell carcinoma)and CNE2 (Poorly differentiated squamous cell carcinoma)in vitro and extract total protein at log phase growth. the total protein of CNE1and CNE2 abstracted by Two dimensional gel electrophoresis and analyzed image by Image Master analytical system after silver staining.MALDI-TOF-MS has been used to analyze protein spots abstracted from the glue to get peptide PMF. Mascot software has been to searched NCBInr data bank to identity the different protein.Lastly,applying cell immunohistochemical staining , we further validate an interesting and differential protein pyruvate kinase sereening from CNE1 and CNE2.RESULTwe get six proteins by Two dimensional gel electrophoresis and appraised by mass spectrogram technology and analyzed by bioinformatics. Cell strain CNE1 specially express proteins: pyruvate kinase,20s Proteasome and beta-actin,Cell strain CNE1 specially express proteins: adenine phosphoribosyltransferase,proteasome beta 6 subunit and platelet-activating factor acetylhydrolase.which were maybe negatively related to metastasis in coloreetal carcinoma.Some of them related to gene transcription, cell cyele, signal transduetion,cell apoptosis,etc,which maybe involved in cell differentiation,Proliferation,invasion , adhesion,metastasis of NPC. from the total protein abstracted cell immunohistochemical staining has been to checked the expression of pyruvate kinase. Using cell immunohistoehemtry ,the expression of pk Protein was diffusely strong positive in CNE1 while the expression was focally positive in CNE2.Conclusionâ… .Proteomic techniques are the ideal methods of quiekly and utilitily screening metastasis associated Proteins of NPC. The experiment obtains protein maps with high resolution and reprodueibility in differently metastatie Potential cell strains, CNE1andCNE2,using 2-DE.â…¡. The 2-DE map of cell strain CNE1 disPlays obvious difference compared with cell strain CNE2 of NPC, There are 6 kinds of Probably metastasis-assoeiated Protein.There are: pyruvate kinase,20s Proteasome, beta-actin,adenine phosphoribosyltransferase,proteasome beta 6 subunit and platelet-activating factor acetylhydrolase.â…¢. Cells by immunohistochemistry using the method of difference in the pyruvate kinase protein further verification, The results showed that: Pyruvate kinase in human nasopharyngeal carcinoma cell line CNE1 stronger than the expression level of human nasopharyngeal carcinoma cell line CNE2. To verify the results of mass spectrometry results are consistentâ…£. This study provide the theory evidences for searching marker to predict NPC metastasis.