人肺腺癌细胞耐顺铂株A549/CDDP的比较蛋白质组学研究

Comparative Proteomic Analysis of Human Lung Adenocarcinoma Cisplatin-resistant Cell Strain A549/CDDP

背景与目的化疗是肺癌综合治疗的重要部分,但是肿瘤细胞耐药常导致化疗失败。本研究采用蛋白组学方法研究人肺腺癌细胞A549对顺铂产生耐药性前后的蛋白表达差异,筛选有价值的靶位。方法以顺铂为诱导药物,人肺腺癌细胞系A549为诱导对象,采用逐步增加剂量与大剂量冲击相结合的方法,诱导建立耐顺铂细胞株A549/CDDP。对A549与A549/CDDP进行蛋白组学研究,即利用双向凝胶电泳(2-DE)分离两组总蛋白后,通过图像分析寻找表达差异的蛋白,对其进行MALDI-TOF质谱分析。结果比较两者2-DE图谱,得到差异蛋白点82个;对其中6个差异蛋白点进行肽质量指纹图分析,鉴定出葡萄糖调节蛋白75、核糖体蛋白S4、线粒体F1-ATP合酶β亚单位、免疫球蛋白重链可变区;以上差异蛋白均在耐药株中过表达,而在对照组细胞中表达下调或不表达。结论所鉴定的差异蛋白将为阐明肺癌细胞对顺铂耐药性产生的机制提供线索,也为筛选具体潜在价值的肺癌化疗靶位提供理论依据。

Background and objective Chemotherapy plays an important role in the comprehensive therapy of lung cancer.However,the drug-resistance often causes the failure of the chemotherapy.The aim of this study is to identify differently expressed protein before and after cisplatin resistance of human lung adenocarcinoma cell A549 by proteomic analysis.Methods Cisplatin-resistant cell strain A549/CDDP was established by combining gradually increasing concentration of cisplatin with large dosage impact.Comparative proteomic analysis of A549 and A549/CDDP were carried out by means of two-dimensional gel electrophoresis.The differentially expressed proteins were detected and identified by MALDI-TOF mass spectrometry.Re-sults Eighty-two differentially expressed proteins were screened by analysis the electrophoretic maps of A549 and A549/CDDP.Six differential proteins were analyzed by peptide mass fingerprinting.Glucose regulating protein 75,ribosomal protein S4,mitochondrial ATP synthase F1 complex beta subunit and immunoglobulin heavy chain variable region were identified.All four differentially expressed proteins were over-expressed in A549/CDDP,whereas low-expressed or no-expressed in A549.Conclusion These differentially expressed proteins give some clues to elucidate the mechanism of lung cancer cell resistant of cisplatin,providing the basis of searching for potential target of chemotherapy of lung cancer.