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Proteomic Analysis of the Anti-Tumor Mechanism of Bufalin

Author: PanSuNa
Tutor: MaHaiLe; FanJinLing
School: Henan University of Science and Technology
Course: Of Food Science
Keywords: bufalin HeLa cell apoptosis Two-dimensional electrophoresis Proteomics
CLC: R285
Type: Master's thesis
Year: 2012
Downloads: 55
Quote: 0
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Abstract


Despite of improvements in treatment of patients with cervical cancer over therecent years, cervical cancer was still one of leading causes of death owing to cancer inwomen. Bufalin is one of strongest toxic compounds isolated from the traditionalChinese medicine Chansu (toad venom). Previous studies showed that bufalinpossessed potent antitumor activites and induced apotosis and differentiation of cancercells. But, the molecular mechanism of bufalin was still not fully clear, and proteomicsresearch of bufalin had not been reported before. In this study, firstly, the cytotoxicityof bufalin in HeLa cells was checked. Then, a comparative proteomics was conductedto find proteins differentially excpressed under bufalin treatment. These possibletarget-related proteins of bufalin were confirmed and the possible mechanism ofbufalin was further studied.Results of MTT assay showed that bufalin treatment for24h inhibited theproliferation of HeLa cells with an IC50value of154±21.5nmol·L-1. Flow cytometricanalysis and Hoechst33342staining analysis indicated that bufalin induced apoptosisin HeLa cells. For protemic study,2D-PAGE was employed to check the proteinexpression profiles of HeLa cells with or without bufalin treatment (24h). Bycomparing the protein expression profiles of bufalin-treated group and control group,11possible target-related proteins of bufalin were found. Among the11proteins,nudix-type motif5, vimentin, hnRNP C1/hnRNP C2variant, hnrpK, hnrpK isoform avariant (two spots are the same protein), heat shock protein27, macrophage-cappingprotein, SELENBP1protein were down-regulated, whereas Ribosomal protein, S-Adenosylmethionine Synthetase2were up-regulated. Consistent with the proteomicstudy results, western blotting assay results showed that heat shock protein27,vimentin, HNRPK were found to be down-regulated in bufalin-treated HeLa cellscompared with control. The possible protein-protein interaction network based ontarget-related proteins of bufalin was constructed using bioinformatic methods. Inonclusion bufalin induced apoptosis in HeLa and the effect of bufalin might be related to its regulation on multiple protein targets.

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