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【目的】寻找并鉴定CNE2细胞在EB病毒感染后表达发生变化的功能蛋白质,为阐明该病毒诱导鼻咽上皮细胞发生生物学变化的作用机制提供线索。【方法】分别从EB病毒感染后的CNE2细胞和未经感染的CNE2细胞中抽提总蛋白质,通过双向电泳分离。运用软件比较分析上述两组的蛋白质表达图谱,寻找表达有差异的蛋白质点。从凝胶上切下差异蛋白质点,通过基质辅助激光解吸/离子化飞行时间质谱(MALDI-TOF-MS)鉴定。【结果】图像分析显示,EB病毒感染后的CNE2细胞和未经感染的CNE2细胞之间存在一些明显差异的蛋白质点。通过MALDI-TOF-MS成功地鉴定了其中4个蛋白质点,EB病毒感染CNE2细胞后蛋白质GRP78和硫氧还蛋白过氧化物酶1表达上调,蛋白质肌动蛋白胞浆组分2和蛋白酶体ι链表达下调。【结论】EB病毒感染鼻咽上皮细胞后引起生物学变化的分子机制可能包括:①诱导细胞大量合成蛋白质以促进增生,同时上调蛋白质GRP78的表达而抗凋亡;②提高细胞内氧化状态而诱导蛋白质硫氧还蛋白过氧化物酶1的表达,并可能通过该蛋白质调控NF-κB和AP-1来影响细胞生长。
【Objective】 The aim of this study was to identify and identify functional proteins that CNE2 cells expressed after EBV infection and provide clues for elucidating the mechanism of the virus induced nasopharyngeal epithelial cells biological changes. 【Method】 Total proteins were extracted from CNE2 cells infected with Epstein-Barr virus and non-infected CNE2 cells respectively and separated by two-dimensional electrophoresis. The software was used to compare and analyze the protein expression profiles of the two groups to find the protein spots with different expression. Differential protein spots were excised from the gel and identified by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS). [Results] The results of image analysis showed that there were some distinct protein spots between EBV-infected CNE2 cells and non-infected CNE2 cells. Four protein spots were successfully identified by MALDI-TOF-MS. The expression of protein GRP78 and thioredoxin peroxidase 1 was up-regulated after EBV infection in CNE2 cells. Protein actin cytosolic fraction 2 and proteasome ι Chain expression down. 【Conclusion】 The molecular mechanism of EBV-induced nasopharyngeal epithelial cell’s biological changes may include: ① inducing large amount of protein synthesis in cells to promote hyperplasia; up-regulating protein GRP78 expression and anti-apoptosis; ② inducing intracellular oxidative status The expression of protein thioredoxin peroxidase 1 may affect the cell growth through the regulation of NF-κB and AP-1 by this protein.