论文部分内容阅读
为筛选表皮生长因子受体(epidermalgrowthfactorreceptor,EGFR)调控的鼻咽癌(nasopharyngealcarcinoma,NPC)细胞的分泌蛋白质,揭示EGFR在NPC发病中的作用机制,采用无血清培养法培养NPC细胞系CNE2,并用转化生长因子(transforminggrowthfactor-α,TGF-α)刺激CNE2细胞24h作为实验组,对照组CNE2细胞不用TGF-α刺激.超滤法脱盐并浓缩两组细胞的培养上清制备分泌蛋白,采用双向凝胶电泳技术(two-dimensionalelectrophoresis,2-DE)分离两组细胞的分泌蛋白,PDquest图像分析软件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(matrix-assistedlaserdesorption/ionizationtimeofflightmassspectrometry,MALDI-TOF-MS)鉴定差异表达蛋白.建立了实验组和对照组CNE2细胞分泌蛋白的2-DE图谱,图像分析识别了22个差异蛋白质点,质谱鉴定了8个非冗余蛋白质,其功能涉及肿瘤细胞侵袭转移、细胞凋亡和增殖,为进一步揭示EGFR在NPC发病中的作用及其机制奠定了基础.
To screen the secretory proteins of nasopharyngeal carcinoma (NPC) cells regulated by epidermal growth factor receptor (EGFR) and to reveal the mechanism of EGFR in the pathogenesis of NPC, NPC cell line CNE2 was cultured in serum-free culture and transformed with CNE2 cells were stimulated by transforming growth factor-α (TGF-α) for 24 h in the experimental group, but not in the control group, TGF-α was stimulated in the control group.The secreted proteins were prepared by desalting and concentrating the culture supernatant of the two groups by using a two- Proteins secreted by two groups of cells were separated by two-dimensional electrophoresis (2-DE) electrophoresis. PDquest image analysis software was used to identify differentially expressed protein spots. Matrix-assisted laser desorption / ionization time-dependent mass spectrometry (MALDI-TOF- MS ) Were used to identify differentially expressed proteins.The 2-DE map of CNE2 cells secreted protein was established in the experimental group and the control group, 22 differentially expressed protein spots were identified by image analysis, and 8 non-redundant proteins were identified by mass spectrometry. Their functions involved in the invasion and metastasis of tumor cells , Apoptosis and proliferation, in order to further reveal the incidence of EGFR in NPC Effects and mechanism of the foundation.