| Objective:In this study,the effects of low-frequency low-intensity ultrasound(LFLIU)combined with curcumin on proliferation,migration,invasion and apoptosis of human glioma cell lines,and the expression of lncrna BLACAT1 and the Enhancer of Zeste Homolog-2(EZH2)subunit of Polycomb Repressive Complex 2(PRC2)were observed.To investigate whether LFLIU combined with curcumin has a significantly better therapeutic effect on glioma than alone,and whether BLACAT1 can accelerate the migration and invasion of human glioma cell line U87 and U251,reduce the ability of apoptosis,thus affecting the progress of glioma,and to study the relationship of BLACAT1/EZH2 axis in glioma cell line U87 and U251.Methods:This study takes human glioma cell lines U87 and U251 as the research object.The action conditions of LFLIU(intensity of 142.0 m W/cm2)and curcumin(concentration of 15 μmol/L)were selected.The cells were divided into control group(Control),curcumin group(C),ultrasound group(U)and curcumin and LFLIU combined group(CU).Firstly,the effects of LFLIU combined with curcumin on the proliferation,migration,invasion and apoptosis of U87 and U251 cells were detected by CCK-8method,Transwell method,and flow cytometry.Secondly,the effect of LFLIU combined with curcumin on the expression of BLACAT1 and EZH2 was detected by q RT-PCR.Western blot was used to detect the effect of LFLIU combined with curcumin on the expression of EZH2,G6 PD,BCL2,and BAX protein.In addition,U87 and U251 cells were transfected.Both cell lines were transfected with short hairpin RNA(sh-RNA)and plasmids(BLACAT1,EZH2)by transient transfection,and then silenced or overexpressed and their transfection efficiency was detected by q RT-PCR and Western Blot.After transfection,U87 and U251 cell lines were used to detect the expression changes of G6 PD,BCL2 and BAX protein by Western Blot method.The potential effects of BLACAT1 on the migration,invasion and apoptosis of U87 and U251 cells were evaluated.Finally,the effect of the BLACAT1/EZH2 regulatory loop on the migration and invasion ability of glioma cells was tested by co-transfection and RIP.Results:1.The CCK-8 and Transwell experimental methods showed that compared with the Control group,the C and U groups treated separately reduced the proliferation,migration and invasion ability of U87 and U251 cells,while compared with the Control group,C and U groups treated separately,The CU combination group significantly reduced the proliferation,migration and invasion ability of U87 and U251 cells,and the difference was statistically significant(P<0.05).2.Flow cytometry showed that compared with the Control group,the C group and U group treated separately promoted the apoptosis of U87 and U251 cells,while compared with the Control group,the C group and U group treated separately,the CU combined group significantly promoted The difference in apoptosis of U87 and U251 cells was statistically significant(P<0.05).3.QRT PCR and Western blot showed that compared with control group,C group and u group,the expression of BLACAT1,EZH2,G6 PD and BCL2 were further down regulated and the expression of Bax was up-regulated in Cu combined group(P < 0.05).4.RT-PCR and Western blot were used to detect the transfection efficiency of silencing BLACAT1 and overexpression of BLACAT1 and EZH2,and the difference was statistically significant(P < 0.05).5.Western blot was used to detect the expression of G6 PD,BCL2 and Bax in U87 and U251 cells after silencing and overexpression of BLACAT1.After silencing BLACAT1,Western blot was used to detect the expression of G6 PD,BCL2 and Bax.The results showed that: compared with the negative control group,the expression of G6 PD and BCL2 decreased significantly,and the expression of Bax increased significantly(P < 0.05).After overexpression of BLACAT1,the results showed that compared with the negative control group,the expression of G6 PD and BCL2 was significantly increased,and the expression of Bax was significantly decreased(P < 0.05).6.Flow cytometry was used to detect the apoptosis of U87 and U251 cells after silencing and overexpression of BLACAT1.The results showed that compared with the negative control group,the apoptosis rate was significantly increased after silencing BLACAT1(P < 0.05).Compared with the negative control group,the apoptosis rate was significantly decreased after overexpression of BLACAT1(P < 0.05).7.BLACAT1 mediates the progression of glioma cells by interacting with EZH2.Compared with the negative control group,the ability of migration and invasion of glioma cells was decreased after silencing BLACAT1,and the inhibition of migration and invasion of U87 and U251 cells was weakened after CO transfection with overexpression of EZH2.Conclusion:Curcumin combined with LFLIU can reduce the expression of BLACAT1,EZH2 and BCL2.BLACAT1 can regulate the expression of BCL2 and other proteins,accelerate the migration and invasion of glioma cells,and reduce the apoptosis of glioma cells.Overexpression of EZH2 can reverse the inhibition of migration and invasion induced by silencing BLACAT1.Curcumin combined with LFLIU can inhibit the apoptosis of glioma cells by inhibiting the BLACAT1/EZH2 axis,which provides a potential therapeutic target for glioma. |
