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中国癌症防治杂志

中国癌症防治杂志 ›› 2014, Vol. 6 ›› Issue (4): 336-341.doi: 10.3969/j.issn.1674-5671.2014.04.04

• 基础研究 • 上一篇    下一篇

10 Gy X 射线照射鼻咽癌移植瘤组织差异表达蛋白质的初步筛选

  

  1. 南宁广西医科大学附属肿瘤医院放疗科；广西壮族自治区妇幼保健院；广西医科大学区域性高发肿瘤早期防治研究教育部重点实验室；西安交通大学第二附属医院肿瘤科； 广西大学亚热带农业生物资源保护与利用国家重点实验室
  • 出版日期:2014-12-25 发布日期:2015-01-12
  • 通讯作者: 朱小东 E-mail:zhuxiaodong83@163.com
  • 基金资助:

    国家自然科学基金资助项目（81360343，30860329）；广西卫生厅科研基金资助项目（重2011076）；广西高等学校重大科研基金资助项目（201101ZD004）；广西重点实验室专项经费资助项目（GK2013 -13-A-02-01）

Screening for proteins differentially expressed between nasopharyngeal carcinoma xenografts sensitive or resistant to 10 Gy X-irradiation using a combination of iTRAQand OrbiTrap MS/MS

  • Online:2014-12-25 Published:2015-01-12

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摘要： 目的 比较不同放射敏感性的鼻咽癌裸鼠移植瘤组织蛋白质表达的差异，筛选出与鼻咽癌放射抗拒性相关的蛋白质。方法 建立人鼻咽低分化鳞癌细胞（CNE-2）及其放射抗拒性细胞（CNE-2R）裸鼠移植瘤模型，予X 射线10 Gy 单次照射移植瘤，提取瘤体组织总蛋白质并酶解成肽段，用iTRAQ 试剂分别标记后再用二维液相色谱分离，在LTQ-Orbitrap高分辨率质谱仪上鉴定并筛选差异表达的蛋白质，并对差异表达的蛋白质进行基因本体论（Gene Ontology，GO）基因功能分类。结果 共鉴定出差异表达的蛋白质61 个，其中上调17 个，下调有44 个，GO 分析显示差异表达的蛋白质主要涉及细胞周期调控、分解代谢、信号通路调节、压力应激反应及氧化还原反应等生物学过程。结论 Annexin2 、PDIA3 、Ubiquitin 、14-3-3δ/ζ、CKAP4 、HSPA8 、NLRC4 等差异表达的蛋白质可能与鼻咽癌放射抗拒性相关。

关键词: 鼻咽肿瘤, 移植肿瘤, 放射抗拒性, 蛋白质, 差异表达

Abstract: Objective To detect proteins differentially expressed between radiosensitive and radioresistant nasopharyngeal carcinoma xenografts exposed to 10 Gy X-irradiation in an effort to identify proteins related to radiotherapy resistance.Methods Xenograft tumors of radiosensitive CNE-2 cells or radioresistant CNE-2R cells were established in nude mice. Proteins were extracted，digested into peptides，and labeled using the iTRAQ kit. Peptide mixtures were separated by 2D-HPLC，then analyzed by LTQ-Orbitrap massspectrometry. Differentially expressed proteins were classified according to gene ontology analysis. Results A total of 61 differentially expressed proteins were identified，of which 17 were up-regulated and 44 were down-regulated. GO analysis showed these proteins to
be involved in key biological processes such as cell cycle regulation，catabolism，signaling pathway regulation，response to pressure tress and redox reactions. Conclusion MudPIT analysis identified the following 7 proteins that may help render nasopharyngeal tumor issue radioresistant: Annexin2，PDIA3，Ubiquitin，CKAP4，HSPA8，14-3-3δ/ζ，and NLRC4.

Key words: Nasopharyngeal neoplasm, Transplantation tumor, Radiation resistance, Proteins, Differentially expressed