Proteomic and bioinformatic analyses of possible target-related proteins of gambogic acid in human breast carcinoma MDA-MB-231 cells.

Abstract

Gambogic acid (GA) is an anticancer agent in phase ‖b clinical trial in China but its mechanism of action has not been fully clarified. The present study was designed to search the possible target-related proteins of GA in cancer cells using proteomic method and establish possible network using bioinformatic analysis. Cytotoxicity and anti-migration effects of GA in MDA-MB-231 cells were checked using MTT assay, flow cytometry, wound migration assay, and chamber migration assay. Possible target-related proteins of GA at early (3 h) and late stage (24 h) of treatment were searched using a proteomic technology, two-dimensional electrophoresis (2-DE). The possible network of GA was established using bioinformatic analysis. The intracellular expression levels of vimentin, keratin 18, and calumenin were determined using Western blotting. GA inhibited cell proliferation and induced cell cycle arrest at G2/M phase and apoptosis in MDA-MB-231 cells. Additionally, GA exhibited anti-migration effects at non-toxic doses. In 2-DE analysis, totally 23 possible GA targeted proteins were found, including those with functions in cytoskeleton and transport, regulation of redox state, metabolism, ubiquitin-proteasome system, transcription and translation, protein transport and modification, and cytokine. Network analysis of these proteins suggested that cytoskeleton-related proteins might play important roles in the effects of GA. Results of Western blotting confirmed the cleavage of vimentin, increase in keratin 18, and decrease in calumenin levels in GA-treated cells. In summary, GA is a multi-target compound and its anti-cancer effects may be based on several target-related proteins such as cytoskeleton-related proteins.

• Authors: Dong Li, Xiao-Yi Song, Qing-Xi Yue, Ya-Jun Cui, Miao Liu, Li-Xing Feng, Wan-Ying Wu, Bao-Hong Jiang, Min Yang, Xiao-Bo Qu, Xuan Liu, De-An Guo
• Journal: Chinese journal of natural medicines (Chin J Nat Med) Vol. 13 Issue 1 Pg. 41-51 (Jan 2015) ISSN: 1875-5364 [Electronic] China
• PMID: 25660287 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

Chemical References

• Antineoplastic Agents
• CALU protein, human
• Calcium-Binding Proteins
• KRT18 protein, human
• Keratin-18
• Vimentin
• Xanthones
• gambogic acid
• Ubiquitin-Specific Proteases

Topics

• Antineoplastic Agents (pharmacokinetics)
• Apoptosis (drug effects)
• Breast Neoplasms (drug therapy, metabolism)
• Calcium-Binding Proteins (genetics)
• Cell Line, Tumor
• Cell Migration Assays
• Cell Migration Inhibition (drug effects)
• Cell Proliferation (drug effects)
• Computational Biology (methods)
• Cytoskeleton (metabolism)
• Electrophoresis, Gel, Two-Dimensional
• Flow Cytometry
• Gene Expression
• Humans
• Keratin-18 (genetics)
• Oxidation-Reduction
• Protein Biosynthesis (drug effects)
• Protein Transport
• Proteomics (methods)
• Transcription, Genetic (drug effects)
• Ubiquitin-Specific Proteases (pharmacokinetics)
• Vimentin (genetics)
• Xanthones (pharmacokinetics)