In the present study we used the two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to compare the protein expression in two cell lines (PC-Cl3 and PC-E1A+raf). PC-Cl3 is a immortalized rat thyroid cell line which in vitro retain biochemical characteristics of differentiated thyroid cell. PC-E1A+raf originates by stable transfection with E1A and v-raf oncogenes of the PC-Cl3 cells, and displays a malignant phenotype. The protein profiles differed between the two cell lines as revealed by visual inspection and by image analysis software. We identified 414 ± 10 (n = 10) spots in PC-Cl3, among these 11 significantly decreased, eight were absent and six increased in 2-D gel obtained with PC-E1A+raf. Five of these spots were analyzed with MALDI-TOF, but only three showed a significant match in the databases used in the bioinformatics analysis (Profound, Mascot, ad MS-fit). In particular, one of the spot showed homology with a GAPDH (glyceraldehydes-3-phosphate dehydrogenase, one with H(+) transporting ATP-synthase and one with PEBP (phoshatidylethanolamine-binding protein). The present work shows that the transfection with E1A and v-raf oncogenes of the PC-Cl3 cells causes the change of protein expression of PC-Cl3. Two out of three of these identified spots (ATP-synthase and GAPDH) seems to be involved in the cell apoptosis.
Proteome analysis of normal and transformed thyroid cells
SCHIAVONE, Roberta;ZILLI, Loredana;ZONNO, Vincenzo;STORELLI, Carlo;VILELLA, Sebastiano
2005-01-01
Abstract
In the present study we used the two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) to compare the protein expression in two cell lines (PC-Cl3 and PC-E1A+raf). PC-Cl3 is a immortalized rat thyroid cell line which in vitro retain biochemical characteristics of differentiated thyroid cell. PC-E1A+raf originates by stable transfection with E1A and v-raf oncogenes of the PC-Cl3 cells, and displays a malignant phenotype. The protein profiles differed between the two cell lines as revealed by visual inspection and by image analysis software. We identified 414 ± 10 (n = 10) spots in PC-Cl3, among these 11 significantly decreased, eight were absent and six increased in 2-D gel obtained with PC-E1A+raf. Five of these spots were analyzed with MALDI-TOF, but only three showed a significant match in the databases used in the bioinformatics analysis (Profound, Mascot, ad MS-fit). In particular, one of the spot showed homology with a GAPDH (glyceraldehydes-3-phosphate dehydrogenase, one with H(+) transporting ATP-synthase and one with PEBP (phoshatidylethanolamine-binding protein). The present work shows that the transfection with E1A and v-raf oncogenes of the PC-Cl3 cells causes the change of protein expression of PC-Cl3. Two out of three of these identified spots (ATP-synthase and GAPDH) seems to be involved in the cell apoptosis.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
