Effects of celecoxib combined with arsenic trioxide on bcr-abl protein and signal transduction in CML primary cells / 白血病·淋巴瘤

ABSTRACT

Objective To explore the effects of celecoxib combined with arsenic trioxide (As2O3) on the mRNA expression,protein expression and protein tyrosine kinase (PTK) activity of bcr-abl fusion gene and its downstream signal transduction in chronic myeloid leukemia (CML) primary cells.Methods The cells were incubated with celecoxib (40 μmol/L) or As2O3 (2 μmol/L) alone and celecoxib (40 μmol/L) combined with As2O3 (2 μmol/L) for 36 hours.The changes of mRNA expression,p210 expression and PTK activity of bcr-abl fusion gene in each group were examined respectively by real time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR),Western blot and PTK activity analysis.The important proteins STAT1,STAT5 and their phosphorylatic proteins p-STAT1 and p-STAT5 and inhibitor of DNA binding 1 (ID1)a common downstream target of oncogenic tyrosine kinase were also analyzed by Western blot.Results The mRNA expressions in control group,celecoxib group,As203 group and celecoxib combined with As2O3 group were (5.97±0.53) ％,(5.74±0.46) ％,(5.94±0.57) ％ and (3.06±0.41) ％ respectively,and the statistical difference was found only between celecoxib combined with As2O3 group and control group (t =28.35,P =0.00).The similar statistic difference was only observed between the two groups from PTK activity tests (t =4.38,P =0.04).Western blot also showed that p210,STAT1,STAT5,p-STAT1,p-STAT5 and ID1 were more extraordinaryly downregulated by celecoxib combined with As2O3 than others treatments.Conclusion Celecoxib combined with As2O3 can downregulate mRNA,p210 expression,PTK activity of bcr-abl fusion gene and inhibit STAT and ID1 signal transduction pathways in a synergistic way.