运用蛋白质组学技术研究EB病毒诱导鼻咽癌细胞CNE2表达变化的蛋白质

Proteins with Altered Expression Induced by Epstein-Barr Virus in Nasopharyngeal Carcinoma CNE2 Cells

【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。

To seek and identify functional protein with altered expression in CNE2 cells after Epstein-Barr virus (EBV) infection and then provide clues to clarify the mechanism of biological changes induced by EBV in nasopharyngeal epithelial cells.The total protein from experimental group-CNE2 cells infected with EBV and control group-CNE2 cells without EBV infection was extracted and then separated by two-dimensional electrophoresis (2-DE) and Coomassie brilliant blue staining respectively. Subsequently, the protein expression patterns of the above two groups were compared and analyzed with software to seek protein spots altered in expression. Then the protein spots were excised from gels and identified by the matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS). Image analysis revealed that there were some obviously different protein spots between experimental group and control group. Four protein spots of them were successfully identified by MALDI-TOF-MS. The result showed that GRP78 and thioredoxin peroxidase 1 were up-regulated while actin cytoplasmic 2 and proteasome iota chain were down-regulated in CNE2 cells after EBV infection. [Conclusion] The results suggest that the molecular mechanism of biological changes in nasopharyngeal epithelial cells induced by EBV might be as follows: (1) EBV might induce cells to synthesize amounts of protein for proliferation, and up-regulate the expression of protein GRP78 to antagonize apoptosis.(2)EBV might increase the oxidation in cells so as to induce the expression of protein 'thioredoxin peroxidase 1', and then regulate the expression of NF-κB and AP-1 via thioredoxin peroxidase 1 to affect cell growth.