论文部分内容阅读
目的:筛选鼻咽癌(nasopharyngeal carcinoma,NPC)转移相关的分泌蛋白质,为阐明NPC转移机制以及筛选NPC转移分子标志物提供实验依据。方法:应用二维凝胶电泳(two-dimensional electrophoresis,2-DE)技术分离一对来自同一亲本,具有不同转移潜能的NPC细胞系5-8F和6-10B细胞的分泌蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和电喷雾-四极杆-串联质谱(ESI-Q-TOFMS/MS)对差异表达的蛋白质点进行鉴定。Western印迹法检测差异分泌蛋白质nm23-H1在两株细胞中的表达水平。结果:建立了5-8F和6-10B分泌蛋白质的2-DE图谱,质谱分析鉴定出14个非冗余的分泌蛋白质,其中Oncop-rotein18等蛋白质在高转移潜能NPC细胞系5-8F中的表达水平高于无转移潜能的NPC细胞系,而nm23-H1等蛋白质在5-8F细胞系中的表达水平低于6-10B细胞系。Western印迹分析证实了nm23-H1在5-8F和6-10B细胞分泌蛋白质中的差异表达水平。结论:所鉴定的14个非冗余的差异分泌蛋白质为研究NPC转移机制以及筛选NPC转移分子标志物提供了实验依据。
OBJECTIVE: To screen secreted proteins associated with nasopharyngeal carcinoma (NPC) metastasis and provide experimental evidence for elucidating the mechanism of NPC metastasis and screening molecular markers of NPC metastasis. METHODS: A pair of secreted proteins of NPC cell lines 5-8F and 6-10B cells with different metastatic potentials from the same parent were separated by two-dimensional electrophoresis (2-DE) The identified protein spots were identified by MALDI-TOF-MS and ESI-Q-TOF MS / MS. Western blotting was used to detect the expression level of differentially secreted protein nm23-H1 in the two cell lines. RESULTS: A 2-DE map of 5-8F and 6-10B secreted proteins was established and 14 non-redundant secreted proteins were identified by mass spectrometry. Proteins such as Oncop-rotein18 were found in high metastatic potential NPC cell lines 5-8F The expression level of NPC cells was higher than that of the non-metastatic potential, while the expression of nm23-H1 and other proteins in 5-8F cell lines was lower than that of 6-10B cell lines. Western blot analysis confirmed the differential expression of nm23-H1 in 5-8F and 6-10B cell-secreted proteins. CONCLUSION: The 14 non-redundant differentially secreted proteins identified provide experimental evidence for the study of NPC metastasis mechanisms and screening of NPC metastatic molecular markers.