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目的观察由于苯并芘(BaP)作用而阻滞于细胞周期特定时相的人胚肺成纤维细胞的蛋白表达改变,探讨苯并芘引起的细胞应激和致癌机制。方法利用流式细胞术对苯并芘作用后的细胞进行分析和分选,采用双向凝胶电泳与相应软件分析苯并芘处理后细胞内蛋白质的表达谱,MALDI-TOF(matrix assisted laser desorption/ionization-time of flight)质谱结合数据库检索鉴定差异表达的蛋白斑点。结果细胞经5μmol/L苯并芘处理后明显阻滞于S期,该时相细胞蛋白质表达的改变,主要包括热休克蛋白HSP70、HSP27,超氧化物歧化酶SOD[Mn],不均一核糖核蛋白H,翻译调控肿瘤蛋白,类肌钙蛋白,α-肌球蛋白,肌动蛋白结合蛋白和纽蛋白等参与氧化应激和肿瘤发生有关的蛋白。结论这些表达改变的蛋白与苯并芘引发的细胞应激和细胞S周期阻滞有关,进一步研究可有助于阐明苯并芘的致癌机理。
OBJECTIVE: To observe the protein expression of human embryo lung fibroblasts (PBMCs) that block in the specific phase of the cell cycle induced by benzopyrene (BaP) and to investigate the cell stress and carcinogenesis induced by benzopyrene. Methods The cells after benzopyrene treatment were analyzed and sorted by flow cytometry. The expression profiles of intracellular proteins after benzopyrene treatment were analyzed by two-dimensional gel electrophoresis and corresponding software, and the MALDI-TOF (matrix assisted laser desorption / ionization-time of flight mass spectrometry combined with database search to identify differentially expressed protein spots. Results The cells treated with 5 μmol / L benzopyrene significantly blocked the cells in S phase. The changes of protein expression in phase cells at this time included heat shock protein HSP70, HSP27, superoxide dismutase SOD [Mn], heterogeneous nuclear Protein H, Translational Regulation of Tumor Proteins, Troponin, α-Myosin, Actin-Binding Protein and Bovine Proteins Involved in Oxidative Stress and Tumorigenesis. Conclusion These altered proteins are related to cell stress induced by benzopyrene and cell cycle arrest. Further study may be helpful to elucidate the carcinogenic mechanism of benzopyrene.