Differential Protein Expressions Between Leukemia HL-60 and Adriamycin-Resistant HL-60 Cell Lines by Comparative Preteomic Analysis

Resistance is a major problem of chemotherapy failure in acute leukemia. Although multi-drug resistance is an important factor, the exact mechanisms of resistance remain to be clarified. With the aim of better understanding the protein involvement in development of resistance mechanisms, a comparative proteomic analysis——two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) was used to compare differential protein expression profiles in adriamycin-resistant acute myeloid leukemia (AML) HL-60/ADR and sensitive HL-60 cell line. Total cellular proteins extracted from HL-60 and adriamycin- resistant HL-60 cells were separated by 2-D gel electrophoresis.Differential expressed proteins were analyzed by mass spectrometry (MALDI-TOF/TOF) and database searching. 16 significantly differentially expressed protein spots were identified, among which 13 protein spots were identified as up-regulated and 3 as down-regulated. The identified proteins were categorized into: (±)metabolic enzymes; (II)proteins related with signal transduction;(III)cell cycle regulators; and(‡W)cellular proliferation and apoptosis proteins. Some of these differential protein expression were confirmed by western blot. In addition, we investigated the expression of distinct proteins in the primary leukemia cells. The results revealed that nucleolar phosphoprotein (B23) over-expressed in the relapsed patients with acute monocytic leukemia (M5). It suggests that B23 might be a useful indicator of prognosis of M5, but the exact role in resistant mechanism is still to be investigated. 2-D DIGE is a useful approach to investigate differential protein expression related treatment resistance.