苯并芘处理HELF阻滞于S期细胞的蛋白质差异表达谱

Differential protein expression profiles of arrested S-phase cells in human embryo lung fibroblast cells caused by benzo(a)pyrene

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 differential protein expression profiles of arrested special phase ceils in human embryo lung fibroblast cells （HELF） caused by benzo（a） pyrene （BaP） and to provide clues to explore molecular mechanism of cellular stress and carcinogenesis induced by BaP. Methods The flow cytometry was used to analysis cell cycle and sort cell after BaP exposure, two- dimensional electroploresis （2-DE） was applied to investigate the protein expression levels of HELF cells, matrix assisted laser desorption/ionization-time of flight mass spectrometry （MALDI-TOF） combined with database search was conducted to identify the differentially expressed proteins. Results S phase arrest was obviously detected with treatment of 5 μmol/L BaP in HELF cells. Statistical analysis showed that 80 proteins spots were altered in sorted S phase HELF cells after BaP treatment and 9 proteins were successfully identified, including HSP70, HSP27, SOD（Mn） and other proteins. Conclusion These affected proteins may be involved in the cellular stress response and cell cycle checkpoint responses to BaP exposure, and may mediate carcinogenesis induced by BaP.