| Objectives:In this paper,we observed the effect of low-intensity ultrasound(LIU)combined with curcumin on EZH2 and its downstream factors expression in human glioma U87 cell line,and explored whether the effect of low-intensity ultrasound and curcumin can synergistically down regulate EZH2 expression.Methods:U87 cells were cultured to logarithmic phase and then treated.The cells were treated with curcumin concentration of 5,10 and 15?mol/L respectively,and treated with ultrasonic irradiation with intensity of 83.4 and 142.0 mW/cm~2respectively for 30s,60s and 90s.The inhibition of cell proliferation was detected with CCK-8 kit,and the OD value at 450nm was analyzed.According to CCK-8factorial analysis results,the most effective curcumin concentration,ultrasonic intensity and action time were set as the following experimental treatment conditions,i.e.combined group(curcumin concentration 15?mol/L,ultrasonic intensity number 83.4 mW/cm2,irradiation time 60s).Western bolt was used to detect the expression of EZH2 and its downstream factors H3K27me,H3K27ac.Results:1.The OD measured by CCK-8 is worth the following conclusions:(1)Low-intensity ultrasound irradiation can inhibit the proliferation of glioma U87 cells.When the ultrasound intensity is 83.4 mW/cm2,its inhibitory capacity and irradiation time and The intens ity was positively correlated(p<0.05).When the ultrasonic intensity is 142.0 mW/cm2,its suppression ability has nothing to do with the irradiation time and intensity(p>0.05).(2)C urcumin inhibits tumor cell growth,and the inhibitory ability is positively correlated with concentration.2.The above results were included in the multivariate analysis of variance,and t he results were as follows:(1)Changes in the effects of ultrasonic irradiation intensity(UL),irradiation time(TIME),and curcumin concentration(CUR)on U87 cell growth inhibition The effects were all statistically significant(p<0.05).C hanges in curcumin concentration had the greatest effect on cell growth inhibition(F=14.13),followed by ultrasound irradiation time(F=7.10),and ultrasound irradiation intensity was the smallest.(2)The change in ultrasonic intensity and the change in irradiation time constitute a first-order interaction,which is statistically significant at the level of?=0.05.(3)There was no statistical significance between changes in ultrasound intensity and changes in curcumin concentration,changes in curcumin concentration,and changes in ultrasound irradiation time.(4)The secondary interaction consisting of three factors is statistically significant at the level of?=0.05.It is believed that there is an interaction between the factors,that is,the three views have a synergistic effect on the proliferation inhibition of U87.(5)According to the variance results,the ultrasound intensity,irradiation time,and curcumin concentration of the ultrasound intensity of83.4 mW/cm2,the action time of 60 or 90 seconds,curcumin concentration of 15 ?mol/L inhibition of glioma cell proliferation maximum.The ultrasound intensity was 83.4 mW/cm2,the action time was 60 or 90 seconds,and the curcumin concentration was 10?mol/L,which had the second lowest inhibitory effect on glioma cell proliferation.Therefore,this condition was selected as the combination group condition.3.Based on multiple comparisons and statistical descriptions of the main effects,the following judgments can be made:(1)Low-intensity ultrasound irradiation can inhibit the proliferation of glioma U87 cells,and the effect of irradiation time 60s is stronger than 30s,and the difference is statistically significant.(p<0.05),there was no significant difference between 90s and 60s of irradiation time(p=0.189).(2)Curcumin inhibits tumor cell growth.The effect of curcumin at a concentration of 15?mol/L is significantly stronger than the concentration of 5?mol/L and 10?mol/L.The difference is statistically significant(p<0.05).The concentration of 5?mol/L and 10?mol/L There was no significant difference between L(p=0.129).4.Both low-intensity ultrasound and curcumin can down-regulate the expression of EZH2,H3K27me,and H3K27ac.And the degree of down-regulation in the combination group was significantly higher than that in the single treatment group(p<0.05).5.H3K27me decreased significantly after inhibiting EZH2,and H3K27ac did not change significantly.Conclusion:Low intensity ultrasound alone or combined with curcumin can inhibit the proliferation of glioma cells and down regulate the expression of EZH2,H3K27me and H3k27ac,and the down-regulation degree of the combined effect is more significant than that of the single treatment group. |
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