JNK Mediated The Reversal Of Multidrug Resistance Protein P-gp Expression By Low-frequency And Intensity Ultrasound Combined With Curcumin In Human Glioma Cells

Objective:The aim of this study was to investigate the effects of low-frequency and intensity ultrasound and curcumin alone or in combination on the proliferation of U87cells and explore whether JNK mediates the effect of multidrug resistance protein P-gp expression in glioma cells.Methods:Human glioma U87 cells were applied and cultured to log phase for experiment.U87 cells were treated with different intensities(3.0,50.4,83.4,142,290,474m W/cm~2)of low-intensity and intensity ultrasound and different concentrations of curcumin(5,10,15,20umol/L).And cell proliferation inhibition was detected by CCK-8assay.The appropriate combinated parameters,the intensity(83.4m W/cm~2)of low-frequency and intensity ultrasound and the concentration(15umol/L)of curcumin were selected by CCK-8 assay.Cell proliferation inhibition by CCK-8 and P-gp and JNK protein expression by western-blot were detected in the groups of the combination of both low-frequency and intensity ultrasound and curcumin,curcumin,and low-frequency and intensity ultrasound.Then the inhibitor of JNK was used to check the expression of P-gp.One-way analysis of variance was performed using SPSS 21.0 statistical software and Graph Pad Prism mapping software to compare whether there was statistical significance among the groups.Results:1.The curcumin alone incubated for 24h could inhibit the proliferation of U87cells and decrease the expression of P-gp and JNK protein.With the increase of concentration,the stronger the effect was,the difference was statistically significant(P<0.05).2.After the low-frequency and intensity ultrasound treatment alone,the proliferation of U87 cells was significantly inhibited when the ultrasound intensity was83.4m W/cm~2with statistical significance(P<0.05).3.Combined application of curcumin and low-frequency and intensity ultrasound can significantly inhibit U87 cell proliferation,reduce the expression of P-gp and JNK protein levels,the intensity is better than the single treatment group,with statistical significance(P<0.05).4.The expression of P-gp decreased significantly after the usage of JNK inhibitor.Conclusion:The proliferation was significant inhibited and P-gp and JNK protein expression was significantly down-regulated compared low-frequency and intensity ultrasound and curcumin combined application with low-frequency and intensity ultrasound and curcumin application alone in U87 cells.The expression of P-gp was down-regulated significantly after JNK was inhibited.