摘要
目的本研究利用蛋白质组学筛选抑癌候选基因CCDC19在鼻咽癌中可能调控蛋白。方法提取CCDC19稳定高表达的鼻咽癌3D8和对照鼻咽癌C6细胞蛋白,并采用双向凝胶电泳(2D-PAGE)技术进行有效分离,图像扫描后获得高分辨率的2D-PAGE图谱。选用差异蛋白质组学技术筛选差异蛋白位点,然后用肽质量指纹谱(peptide mass fingerprinting,PMF)和数据库检索技术给予鉴定。荧光定量PCR和Western blot验证差异基因表达水平。结果质谱分析显示,FASN、CTSD和PGK1在CCDC19过表达的3D8细胞中表达下调,分别为-3.28、-1.64和-6.97倍。荧光定量PCR在mRNA水平验证了FASN、CTSD和PGK1以及Western blot在蛋白水平验证CTSD蛋白在3D8细胞中表达下调。结论 FASN、CTSD和PGK1可能是CCDC19在鼻咽癌中调控的靶基因。
Objective To screen the proteins under regulation by the candidate tumor suppressor gene CCDC19 using proteomics technology in nasopharyngeal carcinoma （NPC）. Method The cellular proteins were extracted from 3D8 NPC cells with CCDC19 overexpression and the control C6 NPC cells. Two-dimensional （2D） gel electrophoresis was employed to compare the protein expression profiles between these two cells, and the differential proteins were identified using peptide mass fingerprinting and database searching. Real-time PCR and Western blotting were used to validate the expression levels of the differential proteins. Results Matrix-assisted laser desorption/time of flight showed that 3 differential proteins, namely FASN, CTSD and PGK1, were down-regulated by -3.28, -1.64, and -6.97 folds, respectively, which were confirmed by real-time PCR and Western blotting. Conclusion FASN, CTSD and PGK1 are probably the target proteins regulated by CCDC19 in NPC.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2012年第8期1127-1130,共4页
Journal of Southern Medical University
基金
国家自然科学基金(81071632)
广东省育苗工程计划(LYM11102)
广州市珠江科技新星计划(2011J2200009)~~
