Sensitivity of human leukemia K562/ADR cells with inhibited JNK signaling pathway to nilotinib

YAN Jing; HE Zhixu; LUAN Zuo

doi:10.3969/j.issn.1002-266X.2017.02.004

Abstract：Objective To observe the influence of inhibiting JNK signaling pathway on the sensitivity of K562 /ADR cells to nilotinib.Methods The inhibitory concentration (IC50 )and resistance index of K562 cells and K562 /ADR cells to nilotinib was measured by CCK-8 assay.K562 /ADR cells were exposed to nilotinib after JNK signaling pathway was in-hibited by 4 μg/mL SP600125.CCK-8 assay was used to detect the IC50 .K562 /ADR cells were divided into the control group,nilotinib group and the combination group.The control group was not treated,nilotinib group was treated with 20μmol/L nilotinib,and the combination group was first treated with 4 μg/mL SP600125 for 2 h,and then treated with 20μmol/L nilotinib.The MDR1 gene was detected by real-time PCR.The P-glycoprotein (P-gp)was detected by Western blotting.Results The resistance index of K562 /ADR cells to nilotinib was 19.58 times higher than that of K562 cells. The IC50 of K562 /ADR cells to nilotinib decreased from (12.53 ±0.11)μmol/L to (6.77 ±0.24)μmol/L after inhibi-tion of JNK signaling pathway (P <0.05).The relative expression of MDR1 mRNA in K562 /ADR cells was 0.967 8 ± 0.006 2,0.513 6 ±0.012 4,and 0.267 1 ±0.008 7 in the control group,nilotinib group and the combination group;the relative expression of P-gp was 1.007 4 ±0.004 1,0.596 1 ±0.013 6,and 0.363 7 ±0.006 9,respectively;and signifi-cant difference was found between every two groups (all P <0.05).Conclusion The sensitivity of human leukemia K562 /ADR cells with inhibited JNK signaling pathway to nilotinib is increased,which is associated with that the inhibition of JNK signaling pathway can decrease the expression of MDR1 gene in K562 /ADR cells.