应用蛋白质组技术筛选人食管癌细胞系9706顺铂耐药相关蛋白

Screening differential proteins involved in esophageal squamous cell line EC9706/CDDP by proteomic approach

目的应用蛋白质组技术筛选人食管癌(esophageal cancer,EC)顺铂耐药相关蛋白。方法采用顺铂(cis-diamminedichloroplatinum,CDDP)浓度递增法、间歇作用方式建立EC9706顺铂耐药细胞系(EC9706/CDDP),应用细胞培养稳定同位素标记(cell culture and stable isotope labeling,SILAC)、质谱(mass spectrometry,MS)筛选和鉴定食管癌耐药相关的蛋白。结果共鉴定出耐药相关的74种差异表达蛋白,其中53种在EC9706/CDDP中表达显著升高和21种表达明显降低。结论筛选的EC9706/CDDP耐药相关蛋白对进一步阐明肿瘤多药耐药分子机制具有重要理论意义和应用价值,并为逆转多药耐药表型的药物设计提供了新的分子靶点。

Objective To screen drug resistance associated proteins implicated in esophageal cancer （EC） by proteomie approach. Methods Cis-diamminedieh[oroplatinum （CDDP） resistant sub-cell line EC9706/CDDP was established by exposure of step-wise increase of CDDP concentration and withdrawal of CDDP followed by cell culture and stable isotope labeling （SILAC） of EC9706/CDDP and its parental sensitive EC9706. Mass spectrometry （MS） and bioinformatics were used to quantify mixed proteome and protein identification. Results A total of 74 differential proteins was identified, including 53 up-regulated and 21 down-regulated proteins. Conclusion EC9706/ CDDP resistant proteins identified in present study lay the foundation for deeper elucidation of molecular mechanisms of multi-drug resistance and provide novel therapeutic targets for drug development.