| Objective: To compare the results of proteomics by before and aftercurcumin treatment and to investigate the multidrug resistance(MDR),mechanism and the reversal mechanism with curcumin.Methods: After thecultured colon carcinoma HCT-8/VCR cells were treated by curcumin，theproteins were extracted. The method of deal with protein samples, proteinquantities,the choice of immobilized pH gradient(IPG) strips, isoelectricfocusing(IEF) conditions were used for establishing two-dimensionalelectrophoresis.Proteins were identified and analyzed by PDQuest8.0software. Proteins were identified by MALDI-TOF-MS/MS andbioinformatics analysis.The interested proteins were further validated byWestern blot and RT-PCR. Results:The best2-DE separation results couldbe done through loading200ug of protein lysate which is directly scrapedoff from the plates, using the non-linear IPG strip and longer isoelectricfocusing runs to eliminate salts, and60,000Volts.Hours.twenty-ninedifferential protein spots were found to be differentially expressed.Nineteenproteins were successfully identified by mass spectrometry and bioinformatics analysis,3protein spots were up-regulated and16proteinspots were down-regulated in curcumin-treated HCT-8/VCR. The proteinsrelated with biological behavior of colon carcinoma cells such as celldivision,apoptosis, apoptosis, glycolysis, MDR, antioxidation action,etc.The expressions of those proteins including GSTP1,PRDX6verified byWestern-blot and RT-PCR were consistent with the results obtained byproteomics.Conclusion:The well-resolved, reproducible2-DE profiles ofmultidrug-resistance reversing of curcumin on human colon carcinomaHCT-8/VCR cells have been primarily established.Nineteen proteins inhuman colon cancer HCT-8/VCR cell and curcumin-treated HCT-8/VCR arefound by proteomic techniques,which may be involved in theVCR—resistance of human colon carcinoma cells. |
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