Establishment of two-dimensional gel electrophoresis profiles of proteome from drug-resistant choriocarcinoma cell line JAR/MTX

Objective:To establish two-dimensional gel electrophoresis profiles of proteome from drug-resistant choriocarcinoma cell line JAR/MTX.Methods:JAR/MTX cells were cultured in RPMI1640 culture medium containing 10% fetal bovine serum,then harvested after digestion by trypsin,and lysed in lysis buffer.After centrifugation,supernatant was separated by isoelectric focusing (IEF),equilibrated and run on SDS polyacrylamide gel electrophoresis (SDS-PAGE),then stained by Coomassie brilliant blue.A well-seperated protein spot was selected from the gel by PDQuest analysis system.Hybrid Quadrupole Time of Flight Mass Spctrometer (Q-TOF-MS),peptide mass fingerprinting (PMF) and database searching were used to identify the protein spot.Results:Clear,well-resolved,reproducible two-dimensional gel electrophoresis profiles of proteome in JAR/MTX cells were obtained.(810±45) protein spots were detected in the two-dimensional gel electrophoresis profiles of proteome on 17 cm IPG strip (pH 3～10) of JAR/MTX cell proteome separated.A match rate of 80.2% of the spots between gels was obtained.One of these proteins was identified by mass spectrometry and database queries.Conclusion:The two-dimensional gel electrophoresis profiles of proteome from JAR/MTX cells are established and lay the foundation of further proteomics research of JAR/MTX cells.