用蛋白质组学技术分析鼻咽癌细胞系放射生物学特性的初步结果

Preliminary study on proteomic technique in radiobiological characteristics in nasopharyngeal carcinoma cell line

目的应用蛋白质组学技术观察不同鼻咽癌细胞系中蛋白表达的差异,以期寻找与鼻咽癌细胞系放射生物学特性相关的蛋白。方法提取细胞总蛋白进行双向凝胶电泳、MALDI-TOF肽质谱指纹图分析、质谱数据的蛋白质库搜寻鉴定。应用Western Blot检测细胞中蛋白质表达。结果差异表达最为显著的蛋白质有9个,与5-8F细胞比较6-10B细胞中4个为下调蛋白,5个为上调蛋白。Western Blot证实CK19和p73在5-8F和6-10B中的表达与蛋白质组结果一致,p73在5-8F中的表达高于6-10B,CK19在6-10B中的表达高于5-8F。结论放射生物学特性不同的鼻咽癌细胞系中存在差异表达蛋白,这可能与其放射生物学特性有关,其中p73可能成为预测的侯选标志物。

Objective To examine the variation of protein expression in nasopharyngeal carcinoma cell lines with different biological characteristics and to identify the radiobiological associated proteins. Methods Biological characteristics of 5-8F and 6-10B were compared by flow cytometry assay after irradiation. The total proteins of 5-8F and 6-10B were separated by immobilized pH gradient（IPG） IEF-SDS twodimensional gel electrophoresis technique. The differentially expressed proteins were cut from the gel and digested into peptides for MALDI-TOF MS and the Q-TOF mass spectrometric analysis. Identification of protein was made through searching in protein sequence database. Protein expressions were examined by western blot and immunohistochemistry method. Results Nine most differentially expressed proteins between 5-8F cell and 6-10B cell were identified, p73 and CK19 expression examined by western blot were conformal with that by proteomic method, p73 expression in 5-8F cell was higher than in 6-10B cell. CK19 expression in 6- 10B cell was higher than in 5-8F cell. Conclusion Differentially expression of proteins exist in nasopharyngeal carcinoma cell lines with different biological characteristics. These proteins may be associated with cell radiobiological characteristic with the p73 as a potential biomarker.