Screen and identification of differential proteins in two early-stage lung adenocarcinoma tissues with and without RASSF1A expression / 第二军医大学学报

ABSTRACT

Objective: To establish a two-dimensional electrophoresis (2-DE) gel map of 2 early-stage lung adenocarcinoma tissues with and without RASSF1A expression, so as to screen and identify differential proteins. Methods: Five early-stage lung adenocarcinoma tissues with RASSF1A expression and 5 without RASSF1A expression were screened out by Western blotting assay. The total soluble proteins of the tissue were extracted and were separated by immobilized pH gradient based two-dimensional gel electrophoresis to set up the 2-DE gel map of the 2 adenocarcinoma tissues. The differentially-expressed proteins were analyzed by PDQuest image analysis software and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS); the protein database was searched to further characterized the differential proteins. Results: A well-reproducible 2-DE gel map of the 2 adenocarcinoma tissues with and without RASSF1A expression was established and 17 differential protein spots were screened out. Nine of 17 differential protein spots were selected for MALDI-TOF-MS study and satisfactory peptide mass fingerprints were obtained for all the 9 spots. Searching of the protein database revealed 5 candidate proteins and they were cytochrome b5, 60S acidic ribosomal protein P2, carbonic anhydrase 1, pyrroline-5-carboxylate reductase 1, and apolipoprotein A-I precursor. Conclusion: We have successfully obtained the 2-DE gel images of 2 early-stage lung adenocarcinoma tissues with and without RASSF1A expression, and from which we have identified 5 differential proteins, which paves a way for studying the signal transduction pathways involving RASSF1A.