Increased expression of epidermal fatty acid binding protein, cofilin, and 14-3-3-sigma (stratifin) detected by two-dimensional gel electrophoresis, mass spectrometry and microsequencing of drug-resistant human adenocarcinoma of the pancreas.
In order to study possible mechanisms leading to chemoresistance in pancreatic adenocarcinoma we examined the global protein expression of pancreatic cancer cells in vitro. We used a cell culture model derived from the adenocarcinoma of the pancreas (EPP85-181P). A classical multidrug-resistant subline, EPP85-181RDB, selected in presence of daunorubicin, and an atypical multidrug-resistant cell variant, EPP85-181RNOV, selected in presence of mitoxantrone, were analyzed using two-dimensional electrophoresis. After staining and image analysis, spots of interest were isolated using preparative two-dimensional electrophoresis and subjected to mass spectrometry and microsequencing. Three proteins, E-FABP, cofilin, and 14-3-3-sigma (stratifin), were overexpressed in chemoresistant cell lines. Cofilin was present in both multidrug in chemoresistant cell lines. Cofilin was present in both multidrug-resistant cell lines. E-FABP and 14-3-3-sigma (stratifin) was found to be overexpressed only in the mitoxantrone-selected atypical multidrug-resistant cell line. The possible significance of these findings is discussed.
Sinha
P
P
Institut für Laboratoriums-medizin und Pathobiochemie, Universitätsklinikum Charité, Medizinsche, Fakultät der Humboldt Universität zu Berlin, Germany. pranav.sinha@charite.de
Hütter
G
G
Köttgen
E
E
Dietel
M
M
Schadendorf
D
D
Lage
H
H
D048948:14-3-3 Proteins; D051339:Actin Depolymerizing Factors; D000230:Adenocarcinoma / Q000378:metabolism*; D014408:Biomarkers, Tumor; D019008:Drug Resistance, Neoplasm; D015180:Electrophoresis, Gel, Two-Dimensional; D005092:Exonucleases; D005095:Exoribonucleases; D006801:Humans; D013058:Mass Spectrometry; D008840:Microfilament Proteins / Q000096:biosynthesis*; D009363:Neoplasm Proteins; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D014176:Protein Biosynthesis; D014407:Tumor Cells, Cultured
D048948:14-3-3 Proteins; D051339:Actin Depolymerizing Factors; D014408:Biomarkers, Tumor; D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D005092:Exonucleases; D005095:Exoribonucleases; C413442:SFN protein, human