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目的:应用蛋白质组学技术比较人卵巢癌细胞株COC1及耐药细胞株COC1/DDP之间的蛋白质差异,寻找与顺铂耐药有关的相关蛋白质。方法:提取细胞株COC1、耐药细胞株COC1/DDP的总蛋白,用双向凝胶电泳(2-DE)的方法获得两细胞株的双向电泳图,选择差异倍数在3倍以上、边界清晰的蛋白质点胶内酶切后,用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)和超高效纳升液相色谱-电喷雾串联质谱(NanoUPLC-ESI-MS/MS)分析鉴定。结果:双向凝胶电泳图谱上检测到1 878个蛋白质点,匹配率为93.6%,差异倍数3倍以上的蛋白质点67个,其中COC1/DDP细胞株高表达有23个,低表达的有44个。选取差异蛋白斑点6个用两种质谱方法初步鉴定了6种蛋白质:dUTP焦磷酸酶、aarF域含蛋白激酶4(ADCK4)、3-磷酸甘油醛脱氢酶(GAPDH)、角蛋白1、角蛋白9、角蛋白10。结论:双向凝胶电泳结合质谱技术可用于肿瘤蛋白质差异的分析,有助于研究肿瘤耐药的机制。
OBJECTIVE: To compare the protein differences between human ovarian cancer cell line COC1 and drug-resistant cell line COC1 / DDP by proteomic techniques and to find out the related proteins related to cisplatin resistance. Methods: The total protein of cell line COC1 and drug-resistant cell line COC1 / DDP was extracted. The two-dimensional electrophoresis image of two cell lines was obtained by two-dimensional gel electrophoresis (2-DE) The proteins were digested with restriction endonucleases and identified by MALDI-TOF-MS and NanoUPLC-ESI-MS / MS. Results: A total of 1 788 protein spots were detected on the two-dimensional gel electrophoresis pattern, with a matching rate of 93.6% and 67 times more than 3-fold difference, of which 23 were highly expressed in COC1 / DDP cell lines and 44 A Six protein spots were selected to identify six proteins using two mass spectrometry methods: dUTP pyrophosphatase, aarF domain containing protein kinase 4 (ADCK4), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), keratin 1, Protein 9, Keratin 10. Conclusion: Two-dimensional gel electrophoresis combined with mass spectrometry can be used for the analysis of tumor protein differences, which is helpful to study the mechanism of tumor drug resistance.