顺铂对人卵巢癌细胞系A2780/DDP和A2780影响的差异表达蛋白质分析

目的:比较不同浓度的顺铂作用人卵巢癌顺铂耐药细胞系A2780/DDP和敏感系A2780 48h的双向凝胶电泳(2-DE)图谱的变化,从整体蛋白质水平对卵巢癌顺铂耐药机制进行初步探讨。方法:MTT法观察顺铂对A2780/DDP和A2780细胞系生长的抑制效果。利用2-DE分离顺铂培养48h的A2780/DDP和A2780细胞总蛋白,Image master图像分析系统分析,从中选取部分分离较好的差异蛋白质点,基质辅助激光解析电离飞行时间质谱分析获取肽质量指纹图谱,PeptIdent软件搜索数据库鉴定蛋白质。结果:获取重复性和分辨率较好的顺铂作用人卵巢癌细胞系A2780/DDP和A2780的2-DE图谱。获取5张肽质量指纹图谱。初步鉴定3种可能影响顺铂抑制A2780/DDP或A2780细胞系生长的蛋白质。结论:建立了5μmol/L和20μmol/L顺铂作用A2780/DDP和A2780的2-DE图谱,并识别鉴定出部分差异表达蛋白质,为阐明卵巢癌顺铂耐药机制提供了实验依据。

Analysis of differentially expressed proteins for the effect of cisplatin on human ovarian cancer cell lines A2780/DDP and A2780

Objective:To compare the two-dimensional polyacrylamide gel electrophoresis(2-DE) maps of human cisplatin-resistant ovarian cell line A2780/DDP with cisplatin-sensitive ovarian cell line A2780 in various dose of cisplatin to explore pathogenesis of cisplatin resistance in ovarian cancer.Methods: The influence of various doses of cisplatin on cell growth in A2780 and A2780/DDP cell lines were detected by MTT.Immobilized pH gradient 2-DE was applied to separate the total cell proteins from A2780 and A2780/DDP cells.Image Master 2-DE analysis software,peptide mass fingerprint based on matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS) and database searching were further applied.Results:2-DE patterns with high resolution and reproducibility of the cell lines were obtained.5 protein spots were incised from sliver staining gel and digested in gel by trypisin.5 maps of peptide mass fingerprints(PMF) were obtained and 3 proteins perhapers had influence on cisplatin-induced cell-growth inhibition.Conclusion:Reference 2-DE maps of A2780 and A2780/DDP proteome treated with 5μmol/L and 20μmol/L cisplatin are established and the significantly different proteins are preliminarily analysed.It provides an experimental basis for the further study of the cisplatin-resistance mechanism in human ovarian cancer.