tailieunhanh - Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin inhibits the proliferation of ARPE-19 cells

The antiproliferative effect of the Hsp90 inhibitor 17-AAG (17-allylamino-17-demethoxygeldanamycin) on human retinal pigment epithelial cells is investigated. Methods: MTT and flow cytometry were used to study the antiproliferative effects of the 17-AAG treatment of ARPE-19 cells. 2D gel electrophoresis (2-DE) and mass spectrometry were applied to detect the altered expression of proteins, which was verified by real-time PCR. Gene Ontology analysis and Ingenuity Pathway Analysis (IPA) were utilized to analyze the signaling pathways, cellular location, function, and network connections of the identified proteins. And SOD assay was. | Yao et al. Journal of Biomedical Science 2010 17 30 http content 17 1 30 a NSC Tha cost of publication In Journal of Blomodlcal Science Is boms by tlM National Science Council Taiwan JOURNAL OF BIOMEDICAL SCIENCE RESEARCH Open Access Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin inhibits the proliferation of ARPE-19 cells Jia-Qi Yao 1 Qing-Huai Liu 21 Xi Chen 1 Qin Yang1 Zhi-Yang Xu2 Fan Hu2 Lin Wang2 and Jian-Min Li 2 Abstract Background The antiproliferative effect of the Hsp90 inhibitor 17-AAG 17-allylamino-17-demethoxygeldanamycin on human retinal pigment epithelial cells is investigated. Methods MTT and flow cytometry were used to study the antiproliferative effects of the 17-AAG treatment of ARPE-19 cells. 2D gel electrophoresis 2-DE and mass spectrometry were applied to detect the altered expression of proteins which was verified by real-time PCR. Gene Ontology analysis and Ingenuity Pathway Analysis IPA were utilized to analyze the signaling pathways cellular location function and network connections of the identified proteins. And SOD assay was employed to confirm the analysis. Results 17-AAG suppressed the proliferation of ARPE-19 cells by inducing cell cycle arrest and apoptosis. Proteomic analysis revealed that the expression of 94 proteins was altered by a factor of more than following exposure to 17-AAG. Of these 94 87 proteins were identified. Real-time PCR results indicated that Hsp90 and Hsp70 which were not identified by proteomic analysis were both upregulated upon 17-AAG treatment. IPA revealed that most of the proteins have functions that are related to oxidative stress as verified by SOD assay while canonical pathway analysis revealed glycolysis gluconeogenesis. Conclusions 17-AAG suppressed the proliferation of ARPE-19 cells by inducing cell cycle arrest and apoptosis and possibly by oxidative stress. Background The pathogenesis of some eye diseases involves the proliferation of retinal pigment epithelial RPE .

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