Effect of Nilotinib on HO-1 Gene Expression and Induction of Apoptosis in K562/A02 Cells

OBJECTIVE To investigate the effect of nilotinib on HO-1 gene expression and induction of apoptosis in K562/A02 cells.METHODS BCR-ABL in K562/A02 cell was detected by Fluorescence in situ hybridization(FISH).K562/A02 cells were treated with AMN107 of different concentrations for 24 h.The expression of BCR-ABL gene on mRNA level was detected by RQ-PCR.Cell proliferation was observed by MTT.Cell apoptosis and cell cycle were inspected by Annexin V/PI double staining method.The expression of HO-1 gene was determined using RT-PCR and Western Blot.RESULTS Ninety-eight percent of K562/A02 cells was detected with BCR-ABL fusion gene by FISH.RQ-PCR results showed that BCR-ABL fusion gene had a dose-dependent relationship with AMN107 in K562/A02 cells treated by AMN107(0,5,10 and 20 μmol·L-1) for 24 h.MTT and Annexin V/PI tests showed that survival rate of treated cells was negatively associated with the concentrations of AMN107,while the apoptosis rate was positively associated.Cell cycle analysis showed that the number of cells in G0/G1 phase decreased sharply and the cell cycle arrested at G2/M checkpoint in AMN107 treated cells.RT-PCR and Western blot tests indicated that the expression of HO-1 gene decreased negatively with increasing concentration of AMN107.CONCLUSION AMN107 can inhibit the expression of BCR-ABL and HO-1,which may be the mechanism underlying AMN107-induced apoptosis of drug-resistant CML cells.The results indicats that HO-1 is a key factor related with the growth of CML cells and the existence of BCR-ABL gene,and it may be a novel target for overcoming resistance of CML.