Dissertation
Dissertation > Medicine, health > Oncology > Department of Otolaryngology tumor > Pharyngeal tumors

Establishment of Protein Profile of Human Nasopharyngeal Carcinoma and Human Nasopharyngeal Epithelial Tissue Purified by Laser Capture Microdissection

Author PengFang
Tutor ChenZhuChu
School Central South University
Course Pathology and Pathophysiology
Keywords LCM nasopharyngeal carcinoma normal nasopharyngeal epithelial tissue proteome profile database
CLC R739.63
Type Master's thesis
Year 2008
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Nasopharyngeal carcinoma (NPC) is one of the most common cancers in southern China and Southeast Asia, which arising from normal nasopharyngeal epithelial tissue (NNET), and it poses one of the most serious public health problems. Although most research about NPC indicates that the development of this cancer may be related to special genetic and environmental factors, the molecular mechanism of NPC carcinogenesis remains poorly understood. High-throughput proteomics has introduced a new approach to cancer research which aims at identifying differential expression proteins associated with the development and progression of cancer, providing new opportunities to uncover biomarkers and therapeutic targets for NPC as well as reveal the molecular mechanism underlying this disease. Therefore, the establishment of protein expression profile of NPC tissue and NNET is helpful to illuminate the physiological function of NNET, as well as provide a better understanding of the molecular mechanisms that participate in the formation of NPC and identify novel NPC biomarkers.Tissue samples may be the most direct and effective way in the reseach of tumor proteomics. However, A major obstacle for the analysis of tumor specimens is tissue heterogeneity, in order to obtain the pure target cells, LCM has emerged as a good choice for purifying cells from tissue.In order to establish the proteomic profile of NPC tissue and NNET, In our study, LCM was used to isolate pure target cell populations from NPC and NNET tissue samples firstly, with the problem of tissue heterogeneity successfully be tackled. And then, four complementary gel-based and non gel-based proteomic approaches were applied in this study: two-dimensional gel electrophoresis in combination with matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (2DE-MALDI-TOF-MS); one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Electrospray ionization-quadrupole time of flight mass spectrometry ((SDS-PAGE+ESI-Q-TOF MS); 2D liquid chromatography plus electrospray ionization ion-trap mass spectrometry(2D LC+ESI-IT MS) and reversed phase liquid chromatography followed by electrospray ionization ion-trap mass spectrometry (RP-LC-IT MS). Results as follow:1. A total of 1037 and 1006 non-redundant proteins were identified respectively with the global proteome profile of NPC and NNET constructed completely, All identified proteins have been classified in terms of their cellular localization, physiological function and physico-chemical property using information from PIGOK、GOA and ExPASy websites.2. two distinct and creditable reference maps of NPC(239 unique proteins in 427 spot) and NNET (227 unique proteins in 419 spots) were also established, the 2DE MS informations were collected by HUP-ML Editor software and imported into the proteomic database system platform, which was built by Apache+ PHP+ Xindice software, Therefore, the detailed information of identified proteins, such as protein name, PI, MW. et al, can be accessed from our website http ://www.xyproteomics.org/en/.In this study, proteome profile of the NPC tissue and NNET purified by laser capture microdissection were initiated as an indispensable prerequisite to make better elucidation of the molecular mechanisms of carcinogenesis in NPC.

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