The Effect Of Combinative Application Of CL And FR On The Human Chronic Myeloid Leukemia K562 Cells

Objective To study how the CL,FR and the combination of the two drugs affect the proliferation and apoptosis of human chronic myeloid leukemia K562 cells,then to explore the potential mechanism.Methods 1.The CCK-8 method was used to evaluate the inhibitory effect of CL,FR and the combination group of different concentrations at 24 h,48h and 72 h. 2.Treating the cells by CL,FR and the combination group for 48 h,then used the Hoechst33342 staining and fluorescence microscope to observe the changes of the opoptosis cell form. 3.Cell cycle analysis was conducted by PI staining and Flow cytometry instrument after the cells were treated by CL,FR and the combination group for 48 h. 4.The opoptosis rate was observed by the Annexin V-FITC/PI staining and Flow cytometry instrument after the cells were treated by CL,FR and the combination group for 48 h. 5.The protein expression of c-myc was detected by Immunocytochemistry after the cells were treated by CL,FR and the combination group.Results 1.CL,FR and the combination group markedly inhibited the proliferation of the K562 cells in dose- and time-dependent manner.with the increase of time and the drug concentration,the antiproliferation effect has gradually increased,and the combined inhibition rate is stronger than the single drug and non-treated group.the differences are statistically significant(P <0.05).2.The phenomenon of nucleus pycnosis fracture has increased after exposure to the drugs for 48 h. Compared to the control and single group,the changes in the combination group is statistically significant. 3.G0/G1 arrest was caused after the cells were treated by the CL,FR and the combination group for 48 h. Comparing with the non-treated and single group,the inhibited cells in the combination group were largely increased(P <0.05). 4.Opoptosis was significantly induced by the CL,FR and the combination group after the cells were treated for 48 h.but the effect of the combination group was more apparent, When comparing with the control and single group(P <0.05). 5.The CL,FR and the combination group obviously reduced the protein expression level of c-myc.the combined group more significant than the monotherapy group.Conclusion The CL,FR and the combination group have an antiproliferation and proapoptotic effect on K562 cells, the antiproliferation effect is dose and time dependent.the CL,FR and the combination group can inhibit K562 cells in G0/G1.the CL,FR and the combination group can decrease the expression of the protein c-myc.the proliferation inhibition,apoptosis, cell cycle arrest and the down-expression of the protein c-myc in the combination group are higher than in the single group.the two drugs have synergistic anti-cancer effect,and the experiment makes a good basis for using the two drugs into clinical therapy of the chronic myeloid leukemia.