蛋白组学分析放射抗拒性鼻咽癌细胞差异表达蛋白
发布时间：2019-02-18 15:12
【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。
[Abstract]:Objective to screen the proteomic differential expression of nasopharyngeal carcinoma cells with different radiosensitivity from the same source, and to explore the mechanism of radioresistance in nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell lines with different radiosensitivity were isolated and analyzed by two-dimensional gel electrophoresis and image analysis software PDQuest. Differential protein spots were identified by LC-MS/MS mass spectrometry and known differential proteins were classified by Gene Oncology Annotation. Some of the selected proteins were verified by Western blot. Results there were 27 candidate proteins in CNE-2 and CNE-2R cells, 11 of which were down-regulated and 16 up-regulated in radiation-resistant CNE-2R cells. Four of them: 4-3-3 蟽, GADD45,PRKDC and HSP27 were confirmed by Western blot method. Conclusion Radio-resistant cells were produced by repeated irradiation of CNE-2 cells, and some mutations occurred at the proteomic level, and these mutations were inherited stably, thus showing the phenotype of radioresistance. By regulating the expression of some proteins associated with radioresistance, it is possible to regulate the radiosensitivity of cells. 4-3-3 蟽, GADD45,PRKDC and HSP27 are potential gene markers for predicting the radiosensitivity of NPC. They may be involved in the production of radioresistance in NPC.
【作者单位】： 西安交通大学医学院第二附属医院肿瘤病院放疗科;
【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)~~
【分类号】：R739.63
[Abstract]:Objective to screen the proteomic differential expression of nasopharyngeal carcinoma cells with different radiosensitivity from the same source, and to explore the mechanism of radioresistance in nasopharyngeal carcinoma. Methods Human nasopharyngeal carcinoma cell lines with different radiosensitivity were isolated and analyzed by two-dimensional gel electrophoresis and image analysis software PDQuest. Differential protein spots were identified by LC-MS/MS mass spectrometry and known differential proteins were classified by Gene Oncology Annotation. Some of the selected proteins were verified by Western blot. Results there were 27 candidate proteins in CNE-2 and CNE-2R cells, 11 of which were down-regulated and 16 up-regulated in radiation-resistant CNE-2R cells. Four of them: 4-3-3 蟽, GADD45,PRKDC and HSP27 were confirmed by Western blot method. Conclusion Radio-resistant cells were produced by repeated irradiation of CNE-2 cells, and some mutations occurred at the proteomic level, and these mutations were inherited stably, thus showing the phenotype of radioresistance. By regulating the expression of some proteins associated with radioresistance, it is possible to regulate the radiosensitivity of cells. 4-3-3 蟽, GADD45,PRKDC and HSP27 are potential gene markers for predicting the radiosensitivity of NPC. They may be involved in the production of radioresistance in NPC.
【作者单位】： 西安交通大学医学院第二附属医院肿瘤病院放疗科;
【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)~~
【分类号】：R739.63
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