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Proteomic and redox-proteomic analysis of berberine-induced cytotoxicity in breast cancer cells
Authors:Chou Hsiu-Chuan  Lu Ying-Chieh  Cheng Chao-Sheng  Chen Yi-Wen  Lyu Ping-Chiang  Lin Cheng-Wen  Timms John F  Chan Hong-Lin
Affiliation:Department of Applied Science, National Hsinchu University of Education, Hsinchu, Taiwan.
Abstract:Berberine is a natural product isolated from herbal plants such as Rhizoma coptidis which has been shown to have anti-neoplastic properties. However, the effects of berberine on the behavior of breast cancers are largely unknown. To determine if berberine might be useful in the treatment of breast cancer and its cytotoxic mechanism, we analyzed the impact of berberine treatment on differential protein expression and redox regulation in human breast cancer cell line MCF-7 using lysine- and cysteine-labeling two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS). This study demonstrated that 96 and 22 protein features were significantly changed in protein expression and thiol reactivity, respectively and revealed that berberine-induced cytotoxicity in breast cancer cells involves dysregulation of protein folding, proteolysis, redox regulation, protein trafficking, cell signaling, electron transport, metabolism and centrosomal structure. Our work shows that this combined proteomic strategy provides a rapid method to study the molecular mechanisms of berberine-induced cytotoxicity in breast cancer cells. The identified targets may be useful for further evaluation as potential targets in breast cancer therapy.
Keywords:1-DE, one-dimensional gel electrophoresis   2-DE, two-dimensional gel electrophoresis   Ab, antibody   BBR, berberine   BSA, bovine serum albumin   CCB, colloidal Coomassie blue   CHAPS, 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate   DCFH-DA, 2,7-dichlorofluorescin diacetate   ddH2O, double deionized water   DIGE, differential gel electrophoresis   DTT, dithiothreitol   EDTA, ethylenediaminetetraacetic acid   FCS, fetal calf serum   MALDI-TOF MS, matrix assisted laser desorption ionization-time of flight mass spectrometry   NP-40, Nonidet P-40   SDS, sodium dodecyl sulfate   TFA, trifluoroacetic acid
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