The Research On Mechanism Of Neferine Enhances The Sensitivity Of STI571 On K562/G01 Cells

OBJECTIVETo observe neferine(nef) neferine enhances the sensitivity of STI571 and the molecular mechanism of reversal multidrug resistance on K562/G01 cells.METHODSCytotoxicity was determined by improved MTT assay.The influence of nef to affect STI571 to induce K562/G01 cells apoptosis was detected by PI staining flow cytometry.The activity of caspase-3 was detected by Caspase 3 Activity Assay Kit.The influence on bcr-abl mRNA and p210Bcr-abl expression was determined by RT-PCR and protein immunoblotting (Western blotting).RESULTS:(1)①Nef 2～64μmol/L,As2O3 10～160μmol/L exhibited an inhibition activity to roliferation in K562/G01 cells. The IC10 of Nef was 8.1±0.16μmol/L and the IC10 of ATO was 10.42±0.64μmol/L.②The IC50 of STI 571 was 0.63±0.04μmol/L in K562 cells and 26.52±1.35μmol/L in K562/G01 cells. Drug resistance activity of K562/G01 cells to STI 571 was 42.1-fold greater than that of K562 cells.③Nef (4μmol/L),Nef (8μmol/L),As2O3 (10μmol/L) decreased IC50 of STI 571 in K562/G01 cells from 29.41±0.48μmol/L to 14.39±0.48μmol/L,8.80±0.25μmol/L和12.75±1.50μmol/L.(2)①The application of single drug of Nef has no effect to induce K562/G01 cells apoptosis (p>0.05). Nef in combination with STI 571 significantly degraded apoptosis than treated by STI571 (p<0.05).②The application of single drug of Nef after 48 h has no effect to inhence caspase-3 activity in K562/G01 cells (p>0.05).Nef in combination with STI571 significantly inhence caspase-3 activity than treated by STI571 (p <0.05). The application of single drug of Nef after 96 h has inhenced caspase-3 activity in K562/G01 cells (p<0.01).(3)①The application of single drug of Nef has no effect on p210Bcr-abl expression in K562/G01 cells (p>0.05). Nef in combination with STI571 significantly degraded p210Bcr-abl expression in K562/G01 cells than treated by STI571 (p<0.05).②The application of single drug of Nef has no effect on bcr/abl mRNA expression in K562/G01 cells (p>0.05). Nef in combination with STI571 significantly degraded bcr-abl mRNA expression in K562/G01 cells than treated by STI571 (p< 0.05).CONCLUSIONSNef has a potency of anticancer to K562/G01 cells. (2) Nef has no effect on bcr/abl mRNA and p210Bcr-abl protein expression but can enhanced the effects of STI571 on down-regulate bcr-abl mRNA and p210Bcr-abl protein expression.Nef is able to potentiate the anticancer function of STI 571.