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目的对结肠癌羟基喜树碱多药耐药细胞(SW1116/HCPT)及其亲代细胞(SW1116)进行蛋白质组学比较研究.探讨肿瘤细胞的多药耐药机制。方法培养羟基喜树碱多药耐药细胞和亲代细胞.提取蛋白质.以固相pH梯度等电聚焦为第一向.十二烷基磺酸钠-聚丙烯酰胺凝胶垂直电泳为第二向进行双向电泳.图像分析软件分析电泳图谱,基质辅助激光解吸电离飞行时间质谱或基质辅助激光解吸电离飞行时间/飞行时间串联质谱鉴定蛋白质。结果发现9个蛋白质点在SW1116/HCPT细胞中表达量发生改变,4个蛋白质点表达量增加,5个蛋白质点表达量减少。质谱鉴定了6个蛋白质点分别为:琥珀酸脱氢酶复合物(亚单位A)、3-磷酸甘油醛脱氢酶、泛素融合降解1相似蛋白、胞核氯离子通道蛋白、β-微管蛋白和ORF蛋白。结论该研究有助于深入理解结肠癌羟基喜树碱多药耐药机制,并有望在新型耐药逆转剂的开发上发挥作用。
Objective To compare proteomics of hydroxycamptothecin-induced multidrug-resistant cell (SW1116 / HCPT) and its parental cell (SW1116) in colon cancer, and to explore the multidrug resistance mechanism of tumor cells. Methods Hydroxycamptothecin multidrug-resistant cells and parental cells were cultured and the proteins were extracted.The isoelectric focusing with solid-phase pH gradient was the first direction.Direct electrophoresis of sodium dodecyl sulfate-polyacrylamide gel was the second Electrophoresis, Matrix Assisted Laser Desorption / Ionization Time of Flight Mass Spectrometry, or Matrix Assisted Laser Desorption / Ionization Time of Flight / Time of Flight Tandem Mass Spectrometry for Protein Identification. The results showed that the expression of 9 protein spots changed in SW1116 / HCPT cells, the expression of 4 protein spots increased and the expression of 5 protein spots decreased. Mass spectrometry identified 6 protein spots as succinate dehydrogenase complex (subunit A), glyceraldehyde-3-phosphate dehydrogenase, ubiquitin fusion degradation 1-like protein, nuclear chloride channel protein, β- Tubulin and ORF proteins. Conclusions This study is helpful to understand the mechanism of multidrug resistance of hydroxycamptothecin in colon cancer and is expected to play a role in the development of new drug resistance reversal agents.