论文部分内容阅读
多药耐药性(multidrug resistance,MDR)是临床肿瘤化疗失败的主要原因之一,定量分析与鉴定食管鳞癌顺铂(cis-diamminedichloroplatinum,CDDP)耐药相关蛋白对阐明食管癌耐药的分子机制具有重要理论意义。本研究采用浓度递增法建立食管癌CDDP耐药细胞系EC9706/CDDP,细胞培养稳定同位素标记(stable isotope labeling with amino acids in cell culture,SILAC)和高效液相-电喷雾串联质谱检测、生物信息学定量分析并鉴定EC9706/CDDP及其母细胞系EC9706的差异蛋白表达谱。EC9706/CDDP细胞呈贴壁性生长但增殖缓慢,多形性、异形性明显,耐药指数为3.23。74种差异表达的蛋白质分子主要包括细胞骨架相关蛋白(20%)、能量代谢相关蛋白(11%)、转录调控及DNA修复相关蛋白(11%)、氧化还原内稳态维持类蛋白(9.5%)、蛋白合成及参与mRNA处理类蛋白(12%)、核糖体结构类蛋白(8.1%)、分子伴侣蛋白(8.1%)、免疫/炎症反应相关蛋白(5.4%)、细胞内转运功能相关蛋白(5.4%)、核组装相关蛋白(2.7%)等。表明食管癌顺铂耐药的发生是多分子参与、多代谢通路失调的复杂过程,差异表达的蛋白分子将有助于深入理解MDR发生的分子机制,并为MDR表型逆转的新型药物设计提供有价值的分子靶点。
Multidrug resistance (MDR) is one of the main reasons for the failure of chemotherapy in clinical tumors. Quantitative analysis and identification of molecules resistant to esophageal cancer by cisplatin (cisplatin-resistant cisplatin) Mechanism has important theoretical significance. In this study, esophageal cancer cell line EC9706 / CDDP, stable isotope labeling with amino acids in cell culture (SILAC) and high performance liquid chromatography-electrospray ionization mass spectrometry (ESI-MS / MS) The differential protein expression profiles of EC9706 / CDDP and its parent cell line EC9706 were quantitatively analyzed and identified. EC9706 / CDDP cells adherent growth but slow proliferation, pleomorphism, heteromorphism obvious, drug resistance index was 3.23.74 kinds of differentially expressed protein molecules include cytoskeleton-related protein (20%), energy metabolism-related protein 11%), transcriptional regulation and DNA repair related proteins (11%), redox homeostasis protein (9.5%), protein synthesis and mRNA processing proteins (12%), ribosomal structural proteins ), Chaperone protein (8.1%), immune / inflammatory response related protein (5.4%), intracellular transport function related protein (5.4%) and nuclear assembly related protein (2.7%). It is indicated that the occurrence of cisplatin resistance in esophageal cancer is a complex process of multi-molecule participation and dysregulation of multiple metabolic pathways. Differentially expressed protein molecules will help to understand the molecular mechanism of MDR and provide a new drug design for the reversal of MDR phenotype Valuable molecular target