Τίτλος:
Pharmacoproteomic study of the natural product Ebenfuran III in DU-145 prostate cancer cells: The quantitative and temporal interrogation of chemically induced cell death at the protein level
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
A naturally occurring benzofuran derivative, Ebenfuran III (Eb III), was investigated for its antiproliferative effects using the DU-145 prostate cell line. Eb III was isolated from Onobrychis ebenoides of the Leguminosae family, a plant endemic in Central and Southern Greece. We have previously reported that Eb III exerts significant cytotoxic effects on certain cancer cell lines. This effect is thought to occur via the isoprenyl moiety at the C-5 position of the molecule. The study aim was to gain a deeper understanding of the pharmacological effect of Eb III on DU-145 cell death at the translational level using a relative quantitative and temporal proteomics approach. Proteins extracted from the cell pellets were subjected to solution phase trypsin proteolysis followed by iTRAQ-labeling. The labeled tryptic peptide extracts were then fractionated using strong cation exchange chromatography and the fractions were analyzed by nanoflow reverse phase ultraperformance liquid chromatography-nanoelectrospray ionization-tandem mass spectrometry analysis using a hybrid QqTOF platform. Using this approach, we compared the expression levels of 1360 proteins analyzed at ≤1% global protein false discovery rate (FDR), commonly present in untreated (control, vehicle only) and Eb III-treated cells at the different exposure time points. Through the iterative use of Ingenuity Pathway Analysis with hierarchical clustering of protein expression patterns, followed by bibliographic research, the temporal regulation of the Calpain-1, ERK2, PAR-4, RAB-7, and Bap31 proteins were identified as potential nodes of multipathway convergence to Eb III induced DU-145 cell death. These proteins were further verified with Western blot analysis. This gel-free, quantitative 2DLC-MS/MS proteomics method effectively captured novel modulated proteins in the DU-145 cell line as a response to Eb III treatment. This approach also provided greater insight to the multifocal and combinatorial signaling pathways implicated in Eb III-induced cell death. © 2013 American Chemical Society.
Συγγραφείς:
Roumeliotis, T.I.
Halabalaki, M.
Alexi, X.
Ankrett, D.
Giannopoulou, E.G.
Skaltsounis, A.-L.
Sayan, B.S.
Alexis, M.N.
Townsend, P.A.
Garbis, S.D.
Περιοδικό:
Journal of Proteome Research
Λέξεις-κλειδιά:
antineoplastic agent; benzofuran derivative; calpain 1; ebenfuran III; natural product; proteinase activated receptor 4; unclassified drug, antiproliferative activity; apoptosis; article; cancer cell; cell death; cell proliferation; cell viability; chemical structure; controlled study; cytotoxicity; human; human cell; IC 50; immunodetection; ion exchange chromatography; legume; molecular weight; Onobrychis ebenoides; priority journal; prostate cancer; protein degradation; protein determination; protein expression; protein secretion; tandem mass spectrometry; ultra performance liquid chromatography, Antineoplastic Agents, Phytogenic; Apoptosis Regulatory Proteins; Benzofurans; Calpain; Cell Death; Cell Line, Tumor; Chromatography, Reverse-Phase; Cluster Analysis; Humans; Male; Membrane Proteins; Mitogen-Activated Protein Kinase 1; Prostatic Neoplasms; Proteins; rab GTP-Binding Proteins; Resorcinols; Tandem Mass Spectrometry, Fabaceae; Onobrychis ebenoides
