蛋白组学分析放射抗拒性鼻咽癌细胞差异表达蛋白  

Proteomic analysis of differentially expressed protein in radioresistance nasopharyngeal carcinoma cells

作　　者：王中卫[1] 王亚利[1] 金迎迎[1] 任宏涛[1] 

机构地区：[1]西安交通大学医学院第二附属医院肿瘤病院放疗科,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2014年第6期810-815,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81141097);陕西省科技发展计划项目(No.2012K13-01-09)~~

摘　　要：目的筛选同一来源放射敏感性不同的鼻咽癌细胞蛋白组学差异表达,探讨鼻咽癌放射抗拒机制。方法建立放射敏感性不同的人鼻咽癌细胞株,采用双向凝胶电泳技术和图像分析软件PDQuest分离和分析不同阶段细胞间的差异蛋白质点,采用LC-MS/MS质谱分析系统鉴定差异蛋白质点,采用Gene Oncology Annotation将已知差异蛋白质进行分类。部分筛选蛋白质采用Western blot进行验证。结果 CNE-2和CNE-2R细胞有差异候选差异蛋白质27个,其中11个蛋白质在放射抗拒CNE-2R细胞中表达下调,16个上调。其中的4个:4-3-3σ、GADD45、PRKDC和HSP27用Western blot的方法得到了进一步确认。结论 CNE-2细胞受射线反复照射产生放射抗拒性细胞,在蛋白组学水平上发生了某些突变,并将这些突变稳定的遗传下来,从而表现出放射抗拒性的表型;通过调控其中与放射抗拒性产生相关的某些蛋白表达,就有可能调控细胞的放射敏感。4-3-3σ、GADD45、PRKDC和HSP27是预测NPC放疗敏感性的潜在基因标志物,它们表达失调可能参与NPC放射抗拒性的产生。Objective To observe the differential gene expression in human nasopharyngeal carcinoma （NPC） cell line with different radiosensitivity by proteomic analysis. Methods We established different radiosensitivity human nasopharyngeal carcinoma cell lines, analyzed and separated the differentially proteins of cells in different stages using two-dimensional gel electrophoresis and image software PDQuest, identified differentially proteins using LC-MS/MS spectrometry system, classified the differences protein using Gene Ontology Annotation. The differentially expressed proteins identified by proteomic approach were further confirmed by Western blot. Results Totally 27 differentially expressed proteins were screened out from different radiosensitivity CNE-2R cell line and its parental CNE-2 cell line, while 16 upregulated proteins in CNE-2R cells and 11 downregulated proteins were found. Among them, 4-3-3σ, GADD45, GRP78 and RKIP proteins were confirmed by Western blot. Conclusion The radioresistant CNE-2R cells have been isolated from the CNE-2 cell line by repeated X-ray irradiation. Their dysregulation may be involved in the development of NPC radioresistance. And 4-3-3σ, GADD45, PRKDC and HSP27 proteins are potential biomarkers for predicting NPC radiosensitivity.

关 键 词：鼻咽癌 蛋白组学 放射抗拒性 

分 类 号：R739.63[医药卫生—肿瘤]