复方斑蝥胶囊血清诱导人肝癌SMMC-7721细胞蛋白质组差异表达分析

目的：考察复方斑蝥胶囊血清对SMMC-7721细胞蛋白质组的影响,从分子水平探讨复方斑蝥胶囊的抗癌作用机制。方法：用血清药理学方法制备复方斑蝥胶囊血清并处理人肝癌SMMC-7721细胞,用双向电泳、图像分析、质谱鉴定等蛋白质组学技术测定复方斑蝥胶囊对癌细胞中有影响的蛋白质。结果：双向电泳分离到SMMC-7721细胞蛋白质组约450个点,其中47个点在对照组和实验组之间有2倍以上量变,质谱鉴定发现实验组出现膜联蛋白A5、热休克70×10³蛋白表达显著上调,真核翻译起始因子5A和硫氧还蛋白过氧化酶2表达显著下调。结论：研究发现了4种与肿瘤的增殖、免疫、凋亡相关的差异表达蛋白质,为进一步研究复方斑蝥胶囊的作用机制提供了线索。血清药理学与蛋白质组学相结合能够有效地展示与药物作用相关的蛋白质,可以作为研究复方药理的一个初步技术平台。

Proteomic analysis of differentially expressed proteins in human hepatoma SMMC-7721 cells induced by Fufang Banmao capsule serum

Objective: To investigate the effects of Fufang Banmao capusle on the proteome of SMMC-7721 cells and discover potential molecular mechanism of anti-cancer at molecular level. Method: SMMC-7721 cells were treated by Fufang Banmao capusle serum prepared with serum pharmacological method; proteomic protocol involving 2-DE, image analysis and mass spectrometry were used to detect the proteins in cells influenced by Fufang Banmao capusle. Result: Approximately 450 protein spots in SMMC-7721 cells were resolved and detected in 2-D gel maps from pH3-10L IEF. 47 protein plots varied over 2-fold quantitively between treated sample and control sample were uncovered. 13 differentially expressed proteins spots were further identified by MALDI-TOF-MS analysis and four of them were successfully identified. Annexin A5, heatshock 70×10³ protein 8 was significantly up-regulated in treated sample compared with control sample, while Eukaryotic translation initiation factor 5A and Peroxiredoxin-2 was significantly down-regulated in treated sample. Conclusion: 4 differently expressed proteins associated with the proliferation, apoptosis, immunity of tumor were detected and they might provide clues for the coming research. The protocol of proteomics combined with serum pharmacological method is an effective platform to research complicated formulas in that it is capable of laying out many proteins associated with Fufang Banmao capusle.