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Comparative Proteomics Studies from the Serum of Nasopharyngeal Carcinoma

Author: LiaoQiuLin
Tutor: DingYanQing
School: First Military Medical University
Course: Pathology
Keywords: Nasopharyngeal carcinoma Serum proteomics Two-dimensional gel electrophoresis Mass Spectrometry
CLC: R739.63
Type: Master's thesis
Year: 2007
Downloads: 128
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Abstract

Nasopharyngeal carcinoma (Nasopharyngeal Carcinoma, NPC) is the primary nasopharyngeal epithelial malignancies, is China's highest incidence of the top ten cancer is the most common head and neck cancer. NPC incidence of occult often been found cervical lymph node and distant metastasis, therefore, early detection and early diagnosis, treatment and prognosis is of great significance, but there is no a fast and effective the NPC early diagnosis means nasopharyngeal it cancer serum proteomic studies on the the NPC pathogenesis of and early diagnosis is important. NPC serum proteomics research the NPC serum protein, research results can be applied to the NPC clinical diagnosis, disease monitoring, efficacy monitoring and prognosis, has important practical value and broad application prospects. NPC serum in the present study is to establish the serum two-dimensional gel electrophoresis as well as to optimize various test conditions based on the use of two-dimensional gel electrophoresis (two-dimensional polyacrylamide gel electrophoresis ,2-DE) MS (mass spectrometry, MS) proteomics research by NPC lymph node metastasis group (n = 37), NPC No lymph node and distant metastasis group (27 cases) and normal control group (n = 42) serum 2-DE map, to identify differentially expressed proteins, protein identification NPC and NPC metastasis-related protein. And NPC proteins identified the validation of serological and immunohistochemical verification. The specific results are as follows: 1. In experimental serum two-dimensional gel electrophoresis, serum samples in the two-dimensional gel electrophoresis of serum collection, preservation, removal of high-abundance proteins in serum electrophoresis conditions, the sample volume, IPG dry strips selection and the buffer The liquid preparation conditions and technical methods to be adjusted to establish a more stable and better reproducibility of serum two-dimensional gel electrophoresis, 2-D map resolution is improved to some extent, effective separation of protein spots increased significantly, many low-abundance proteins were separated. 2. The NPC serum comparative proteomics research, Patterns of 29 differential protein spots successfully identified 23 proteins by mass spectrometry. Compared with the normal group, transferrin, zinc finger protein 544, thyroid hormone binding protein, low expression of nm23-H1 protein and FAD synthetase in the two groups of patients with nasopharyngeal carcinoma; 12 - lipoxygenase, serum amyloid Al precursor, cytochrome P450, sICAM-1, cathepsin G and histone lysine-specific demethylase 1 and other highly expressed in the two groups of nasopharyngeal carcinoma, of which 12 - lipoxygenase, sICAM-1 cathepsin G and histone lysine-specific demethylase 1 increased expression of heat shock protein 70 expression only in nasopharyngeal carcinoma metastasis in nasopharyngeal carcinoma metastasis group than that without metastasis. 3. Verify the identification of the NPC protein experiments, sICAM-1, SAA and HSP70 these three proteins in the the nasopharyngeal group and the normal control group, lymph node metastasis and without lymph node metastasis were significant differences found by the ELISA assay . And these three proteins are closely related to the staging of nasopharyngeal carcinoma, with the improvement of the clinical stage, serum levels and the positive rate was gradually increased. Bivariate correlation analysis showed that these three proteins in the serum of patients with nasopharyngeal carcinoma relevant to all. The immunohistochemistry validation experiments showed that 57 patients with nasopharyngeal carcinoma tissue ICAM-1 and HSP70 expression rates were 75.4% and 63.2%, and ICAM-1 and HSP70 and the clinical stage of NPC, also found that ICAM-1 and HSP70 expression rate was significantly higher than in the lymph node metastasis of nasopharyngeal carcinoma nasopharyngeal carcinoma without metastasis, ICAM-1 and HSP70 and metastasis of nasopharyngeal carcinoma has a certain relevance. ICAM-1 and HSP70 can be used as a reference to determine the clinical stage of nasopharyngeal carcinoma and prognosis indicators. This study established the serum two-dimensional gel electrophoresis, for further analysis, identification and finding disease-related protein to lay a certain foundation; 23 NPC serum protein were identified and validated protein. The results suggest that they may be related to the occurrence and development of NPC, and is expected to be potential markers of nasopharyngeal carcinoma early diagnosis and treatment.

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