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[复方斑蝥胶囊含药血清诱导人肝癌SMMC-7721细胞差异表达蛋白质的蛋白质组学分析]

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

作者信息

Cao Yong-Yan, Wang Meng, Huang Kai-Shun, Yang Qiang, Ding Min

机构信息

Department of Laboratory Medicine, Key Laboratory of Clinical Laboratory Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2007 May;32(9):831-4.

Abstract

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 x 10(3) 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.

摘要

目的

研究复方斑蝥胶囊对SMMC-7721细胞蛋白质组的影响,从分子水平探讨其抗癌的潜在分子机制。

方法

采用血清药理学方法制备复方斑蝥胶囊含药血清处理SMMC-7721细胞;运用包括二维电泳(2-DE)、图像分析和质谱分析在内的蛋白质组学技术,检测受复方斑蝥胶囊影响的细胞中的蛋白质。

结果

在pH3-10L等电聚焦的二维凝胶图谱中,SMMC-7721细胞中约450个蛋白质点得到分辨和检测。发现处理样品与对照样品之间有47个蛋白质点的定量变化超过2倍。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析进一步鉴定了13个差异表达的蛋白质点,其中4个被成功鉴定。与对照样品相比,处理样品中膜联蛋白A5、热休克蛋白70×10³ 8显著上调,而真核翻译起始因子5A和过氧化物酶体增殖物激活受体2在处理样品中显著下调。

结论

检测到4种与肿瘤增殖、凋亡、免疫相关的差异表达蛋白质,可能为后续研究提供线索。蛋白质组学技术与血清药理学方法相结合的方案是研究复杂方剂的有效平台,因为它能够展示许多与复方斑蝥胶囊相关的蛋白质。

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