Dissertation > Excellent graduate degree dissertation topics show

Comparative Proteomic Analysis on Radioresistance of Nasasopharyngeal Carcinoma Cell

Author: HuangShiZuo
Tutor: ZhuXiaoDong
School: Guangxi Medical University
Course: Oncology
Keywords: Nasopharyngeal Radiation resistance Proteomics Two-dimensional gel electrophoresis Matrix-assisted laser desorption / ionization time-of-flight tandem mass spectrometry
CLC: R739.63
Type: Master's thesis
Year: 2011
Downloads: 59
Quote: 0
Read: Download Dissertation

Abstract

Study Background and Objective】 NPC (nasopharyngeal carcinoma, NPC) is one of the common malignant tumors in China, the southern region is particularly high incidence of radiotherapy is considered to be the preferred treatment. However, some patients in clinical radiotherapy after local residue with local recurrence, tumor cell radiation resistance is one of the important reasons. At home and abroad at the gene level of tumor radiation resistance mechanisms conducted a lot of research, cell cycle regulation, apoptosis and DNA damage repair genes and proteins related to, but not yet found in nasopharyngeal expression can good predictor radiation oncology resisted and prognosis of gene or protein, It is speculated that the main reason may be the abundance of mRNA expression of its corresponding protein expression levels inconsistencies caused. This information, the occurrence of nasopharyngeal radiation resistance mechanisms change caused not by a single gene or protein, the result of the interaction of a molecular network but the multi-gene or protein. Therefore, the application of proteomics technology offers the possibility to radiotherapy for nasopharyngeal carcinoma resist further explore the mechanism from the protein expression levels. Currently, nasopharyngeal proteomics research has focused on the nasopharyngeal early diagnosis and screening tumor markers. Tumor cells cultured in vitro study, clinical tissue samples induced tissue heterogeneity can be avoided, the complexity of influencing factors, the loss of positive results many shortcomings. Therefore, a gradual transition from tumor cell lines in vitro clinical become one of the main methods of today's study radiation resistance mechanisms. The purpose of this study is to the application of proteomics technology to find with radiotherapy for nasopharyngeal carcinoma to resist occurrence associated candidate proteins, through the establishment and optimization of cell protein sample preparation methods, stable radiation nasopharyngeal carcinoma cell line resistant to its parental cell comparative proteomics analysis, in order to find and radiation resistance associated differentially expressed proteins. Intended to elucidate the molecular mechanisms of nasopharyngeal carcinoma resist initially in protein expression levels, and to lay the foundation for the subsequent candidate protein biology, functional verification, and animal experiments, individual radiosensitivity of clinical predictors of nasopharyngeal cancer patients in the future and implement individualized radiotherapy new ideas. [Methods] 1, three methods were used on nasopharyngeal carcinoma cells CNE-2 total protein extraction and two-dimensional electrophoresis analysis, the establishment of stability and repeatability sexy cell protein sample preparation method. 2, cultured in vitro with stable radiation resisted cell line CNE-2R and its parental cell line CNE2 were extracted in the logarithmic growth phase CNE-2R, total protein of CNE2 cells. Using two-dimensional gel electrophoresis (2-DE) first to immobilized pH gradient electrophoresis second SDS-polyacrylamide acrylamide gel electrophoresis. Isolated cells CNE-2R, CNE-2 total protein. 3, electrophoresis after silver staining, the application of the ImageMaster 2-DE Platinum 5.0 software for gel analysis, a difference of more than 2 times the protein expression levels of protein spots, cut out the different protein spots on the gel, parallel decolorization, enzymolysis , extraction, and lay the groundwork for subsequent mass spectrometry analysis. 4, using matrix-assisted laser desorption / ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS) differences in protein spots selected for protein identification analysis. Mascot protein database search, protein identification, through a manual search of the literature databases, in-depth analysis of the function of the protein. [Results] 1 Universal lysates lyse the cells in the sample before isoelectric focusing Add Destreak reagents, you can get a good stability, image clear, high-resolution two-dimensional electrophoresis map. By two-dimensional gel electrophoresis, to get a better separation of the cell lines with stable radiation resisted CNE-2R and its parental cell line CNE-2 total protein. ImageMaster 2-DE Platinum 5.0 software to filter out the radio-resistant cell line CNE-2R and the parental cell line CNE-2 between the differences in more than 2 times the difference in protein spots 32. Screened 32 differentially expressed protein spots further line identified by mass spectrometry, 11 protein spots were successfully identified and further line bioinformatics analysis found that most of the protein signal transduction related proteins, chaperones, cytoskeletal components, mainly in terms of the biological function and apoptosis regulation, cell cycle regulation, RNA transcription, signal transduction, cytoskeleton composition and radiation stress response, and so on. These proteins can be used as a potential target for radiation therapy, an effective way to improve tumor radiosensitivity, in-depth study of the value, to provide new ideas and methods of radiation therapy for nasopharyngeal carcinoma. [Conclusions] 2-DE combined with MALDI-TOF-MS technology to identify and identification of resistance to radiotherapy proteins differentially expressed between the cell and its parental cell line. The study found that 11 differentially expressed proteins, whose functions involve a variety of physiological metabolism and regulation process, suggesting nasopharyngeal radiation resistant to multiple proteins together, which may be involved in radiotherapy for nasopharyngeal carcinoma occurrence of resistance.

Related Dissertations

CLC: > Medicine, health > Oncology > Department of Otolaryngology tumor > Pharyngeal tumors
© 2012 www.DissertationTopic.Net Mobile