| Objective: STGC3 was a candidate suppressor gene associated with nasopharyngeal carcinoma (GenBank accession No:AY078383). This study was designed to reveal the preliminary function of STGC3 to NPC cell line (CNE2).Methods: The total proteins of CNE2 , pcDNA3.1(+)/CNE2 and pcDNA3.1(+)/STGC3 /CNE2 were seperated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). After 2-DE, gels were silver staining and Coomassie brilliant blue staining, the differential expression protein spots were analyzed with using ImageMaster analysis software, then identified by peptide mass fmgerprint(PMF) based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. Results: The results showed that the good 2-DE pattern including high resolution and reproducibility was obtained, the protein spots in CNE2, pcDNA3.1(+)/CNE2, pcDNA3.1(+)/STGC3 /CNE2 were 675±27, 660±23 and 662±18 respectively. We compared the 2-DE protein patterns of the gels of the three cell lines, 23 differential protein spots were showed in transfected cell lines, 15 of them realized up-regulation while 8 of them realized down-regulation. The different gel spots were excised and digested in situ, measured with MAIDI-TOF-MS and searched in related database with Mascot software. 18 proteins were preliminarily identified. These proteins were related to cell metastasis, transcription regulation, signaling pathway etc. There was a significant difference at protein level between CNE2 and pcDNA3.1(+)/STGC3 /CNE2 cells. Conclusion:1 .The protein expression profiles of NPC cell lines(CNE2) can be affected by the overexpression of STGC3 gene.2. Up-regulated expression of CyclophilinA ^ Alpha enolase> nM23 protein and down-regulated expression of hnRNP F ^ Thioredoxin were related to cell metastasis, transcription regulation, signaling pathway etc.Objective: Our previous study indicated:There were a delay in tumor formation and a dramatic reduction in tumor size when the cells with overexpression of STGC3 were injected into nude mice, and the tumor size in female nude mice was highly smaller than that in male. This paper was designed to investigate the effect of estrogen on the STGC3-transfected CNE2 cells and its related molecular mechanisms. Methods:The proliferative capacity of the CNE2 cell and pcDNA3.1(+)/STGC3 /CNE2 cell in the culture medium with 17-|3-E2 was evaluated by the means of the microculture tetrazolium assay (MTT). A series of methods, including immobilized pH gradient-two dimensional polyacrylamide gel electrophoresis, silver staining, Imaging Master software analysis, peptide mass fingerpringting based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and Mascot database searching, were used to separate and identify the differentially expressed protein induced by estrogen on STGC3 -transfected CNE2 cells.Results: (1) MTT assay showed that the proliferation of CNE2 cell and pcDNA3.1(+)/STGC3/CNE2 cell were significantly inhibited by 17-P-E2 (0.001-1 Oumol/L) in concentration-dependent manner, the inhibitive rate were 6.0%-35.7% and 18.9%-57.7% respectively. The inhibitive effect of 17-p-E2 on pcDNA3.1(+)/STGC3/CNE2 cell was more obvious than on CNE2 cell. (2) After 2-DE, There were 662 ±38 ? 682±43 and 645±56 , 657±61 spots in CNE2. pcDNA3.1(+)/STGC3/CNE2 and CNE2 , pcDNA3.1(+)/STGC3/CNE2cell line dealed with 17-P-E2. The 28 differential protein spots were identified by Imaging...
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