Back to Search
Start Over
Proteomic Analysis Revealed the Important Role of Vimentin in Human Cervical Carcinoma HeLa Cells Treated With Gambogic Acid
- Source :
- Molecular & Cellular Proteomics. 15:26-44
- Publication Year :
- 2016
- Publisher :
- Elsevier BV, 2016.
-
Abstract
- Gambogic acid (GA) is an anticancer agent in phase IIb clinical trial in China. In HeLa cells, GA inhibited cell proliferation, induced cell cycle arrest at G2/M phase and apoptosis, as showed by results of MTT assay and flow cytometric analysis. Possible target-related proteins of GA were searched using comparative proteomic analysis (2-DE) and nine proteins at early (3 h) stage together with nine proteins at late (24 h) stage were found. Vimentin was the only target-related protein found at both early and late stage. Results of both 2-DE analysis and Western blotting assay suggested cleavage of vimentin induced by GA. MS/MS analysis of cleaved vimentin peptides indicated possible cleavage sites of vimentin at or near ser51 and glu425. Results of targeted proteomic analysis showed that GA induced change in phosphorylation state of the vimentin head domain (aa51-64). Caspase inhibitors could not abrogate GA-induced cleavage of vimentin. Over-expression of vimentin ameliorated cytotoxicity of GA in HeLa cells. The GA-activated signal transduction, from p38 MAPK, heat shock protein 27 (HSP27), vimentin, dysfunction of cytoskeleton, to cell death, was predicted and then confirmed. Results of animal study showed that GA treatment inhibited tumor growth in HeLa tumor-bearing mice and cleavage of vimentin could be observed in tumor xenografts of GA-treated animals. Results of immunohistochemical staining also showed down-regulated vimentin level in tumor xenografts of GA-treated animals. Furthermore, compared with cytotoxicity of GA in HeLa cells, cytotoxicity of GA in MCF-7 cells with low level of vimentin was weaker whereas cytotoxicity of GA in MG-63 cells with high level of vimentin was stronger. These results indicated the important role of vimentin in the cytotoxicity of GA. The effects of GA on vimentin and other epithelial-to-mesenchymal transition (EMT) markers provided suggestion for better usage of GA in clinic.
- Subjects :
- Proteomics
0301 basic medicine
Time Factors
Proteome
Cell Survival
Xanthones
Blotting, Western
Mice, Nude
Uterine Cervical Neoplasms
Apoptosis
Vimentin
macromolecular substances
Bioinformatics
Biochemistry
Analytical Chemistry
HeLa
03 medical and health sciences
chemistry.chemical_compound
Tandem Mass Spectrometry
Cell Line, Tumor
Animals
Humans
Electrophoresis, Gel, Two-Dimensional
MTT assay
Cytotoxicity
Molecular Biology
Cell Proliferation
Microscopy, Confocal
biology
Cell growth
Research
biology.organism_classification
Xenograft Model Antitumor Assays
Molecular biology
Tumor Burden
G2 Phase Cell Cycle Checkpoints
030104 developmental biology
chemistry
Cell culture
MCF-7 Cells
biology.protein
Female
RNA Interference
Gambogic acid
HeLa Cells
Subjects
Details
- ISSN :
- 15359476
- Volume :
- 15
- Database :
- OpenAIRE
- Journal :
- Molecular & Cellular Proteomics
- Accession number :
- edsair.doi.dedup.....db71bba0720573dea5438b767276d8fb