Analysis of protein expression in retinoic acid-induced HL60 cells by modified two--dimensional electrophoresis / 重庆医科大学学报

ABSTRACT

Objective:To establish the granulocyte-differentiation model of the HL60 cells which are treated with All-trans retinoic acid(ATRA),and to use the modified two-dimensional electrophoresis conditions to analyze the differences of protein expression between treated and untreated HL60 cells.Methods:HL60 cells were induced through treatment with All-trans retinoic acid(ATRA).For selection of the appropriate drug concentration and induction time,MTT and flow cytometry are used to detect the HL60cell proliferation and the expression of differentiation antigens CD11b respectively.Cellular chemical staining was used for the verification of the differentiation of the treated HL60 cells.The protein of HL60 cell lines could be separated by modified two-dimensional electrophoresis(2-DE).PDQuest software was used to analyze the different protein expression between treated and untreated HL60 cells.The protein was analyzed by matrix-assisted laser desorption -time of flight mass spectrometry(MALDI-TOF).Results:ATRA could inhibit HL60 cell proliferation,and with the increase in drug concentration,the effect of inhibiting was more significant.Treated with 2? M ATRA for five days,there were more than 90% of HL60 cells expressing antigenCD11b.Cellular chemical staining also showed that ATRA could induce HL60 cells to granulocyte cells.By the analysis of modified 2-DE and PDQuest software,25 protein spots was detected in untreated cells,while 15 protein spots was promoted Some of them were oncogene protein and suppressor gene protein,while some of others are involved in apoptosis.Conclusion:ATRA could induce HL60 cells to granulocyte cells in selected drug concentration and induction time.Using the modified two-dimensional electrophoresis conditions,different protein expression can be found from the traditional two-dimensional electrophoresis.