Objective: To obtain and identify the differential proteome of apoptosis induced by realgar (tetra-arsenic tetra-sulfide, As(4)S(4)) in retinoid acid (RA) resistant human acute promyelocytic leukemia (APL) cell line NB4-R1 cells.
Methods: The comparative proteomic expressive profiles of NB4-R1 cells treated with and without As(4)S(4) were obtained with the high-resolution two-dimensional electrophoresis system. After the analysis of ImageMaster(TM) 2D Platinum software combining artificial comparison. The differential expression proteins were screened and performed in-gel digestion and extraction of peptides, applied mass spectrometry MALDI-TOF-MS, MALDI-TOF/TOF, UPLC-MS/MS and bioinformatics to identify differential expressive proteins.
Results: Twenty-two spots with more than 2-fold (≥ 2 or ≤ 0.5) expression changes were identified and 21 known proteins obtained, including 5 down-regulated (SET/I2PP2A, RPP2, PCBP1, EIF4H-1 and ANP32A/I1PP2A) after exposed to As(4)S(4), 2 up-regulated (HSP27 and HMGB1) after exposed for 24 h, and 14 up-regulated (PHB, ERP29, DNAJC8, PSMB4, ACTB, RPP0, RhoGDI2, alpha-tubulin, transcription factor, RBM15/OTT1, eIF5A1 and H2B1M et al.) after exposed for 48 h.
Conclusion: It is the first time to successfully obtain and identify the global proteome of apoptosis induced by As(4)S(4) in RA-resistant cells. Among them, expressional and functional regulation of target proteins SET, RPP2 and PHB might be the potential novel therapeutic target for RA-resistant APL.