Effects Of Neferine With STI571on The Expression Of Hsp90and Inhibitor Of Apoptosis Protein In CML Cells

OBJECTIVE:To further explore the mechanism of neferine enhance STI571sensitivity on the chronic myelogenous leukemia (CML) cells, investigate the effects of Neferine with STI571on bcr/abl、Jak-stat、Hsp90and inhibitor of apoptosis protein(survivin and livin) in CML cells.METHODS:Gene expression and protein levels was detected by Reverse transcriptase quantitative real-time fluorescence polymerase chain reaction (RT-qPCR) and Western blotting respectively.RESULTS:(1) Nef and STI571can both downregulation the expression of bcr/abl (P<0.01).(2) Compared with control group, Stat5A mRNA levels were significantly lower in each drug treatment group (P <0.05).(3) Except Nef4μM group, there are significant differences in Hsp90AA1and Hsp90AB1gene expression of CML primary cells after drug treatment compared with the control group (P<0.01); Compared with STI571alone group, Hsp90AA1and Hsp90ABl gene expression decreased significantly in Nef4μM, Nef8μM combined STI571group (P <0.01).(4) Compared with control group or STI571alone group, the protein levels of Hsp90were significantly decreased in Nef4μM, Nef8μM combined STI571group,(P<0.05).(5) Both STI571and Nef can lower Survivin mRNA levels (P<0.01), and a greater degree of reduction was found when combined Nef4μM or Nef8μM. with STI571.(6) Compared to control group, the level of livin is significant lower in Nef4μM or Nef8μM combined STI571group (P<0.05); Compared to STI571alone group, the level of livin is significant lower in Nef8μM combined STI571group (P<0.05)。CONCLUSIONS:(1) Nef and STI571can reduce level of bcr/abl and Stat5A mRNA on CML cells.(2) Nef and STI571can downregulation gene expression of Hsp90, and further decreased compared to STI571alone When combined Nef with STI571; Compared to control group or STI571alone group, Hsp90protein level reduced in Nef4μM, Nef8μM combined STI571group.(3) Nef and STI571can reduce mRNA levels of survivin, When combined Nef with STI571, gene expression of survivin further decreased compared to STI571alone.(4) The expression of livin gene is down regulated when combined Nef4μM or Nef8μM with STI571.