The Effect Of Ursolic Acid On Autophagy-related Proteins In Human Lung Adenocarcinoma A549 Cells

Purpose: To study the inhibitory effect of ursolic acid on human lung adenocarcinoma A549 cells and the expression of autophagy related proteins LC3,beclin-1 and P62,so as to provide experimental basis for clinical application of ursolic acid to improve the treatment of non-small cell lung cancer.Material and method: Different concentrations of ursolic acid(10,20,30,40,50 ? mol / l)were used to culture human lung adenocarcinoma A549 cells.At 12 h,24h,48 h,CCK8 method was used to detect the effect of ursolic acid on the proliferation of human lung adenocarcinoma A549 cells,and to determine the optimal concentration and time of administration.Human lung adenocarcinoma A549 cells were divided into three groups,namely blank control group,ursolic acid group,ursolic acid and inhibitor group,and then fully cultured After 24 hours of culture,the morphological changes of human lung adenocarcinoma A549 cells were observed by inverted microscope;the expression of autophagy related genes LC3,beclin-1 and P62 were detected by RT-PCR;the expression of autophagy related proteins LC3,beclin-1 and P62 were detected by WB.Results: 1.The results of CCK8 showed that ursolic acid at different concentrations inhibited the proliferation of A549 cells in 12 h,24h and 48 h,and the inhibition was dose-dependent and time-dependent,and compared with ursolic acid group,3-m A improved the inhibition of ursolic acid on A549 cells(P < 0.01).2.The morphological changes of human lung adenocarcinoma A549 cells were observed under inverted microscope.In the blank control group,human lung adenocarcinoma A549 cells grew close to the wall,in good condition,and closely connected with each other.The cells were polygonal or short fusiform,with stretched body,consistent morphology and clear boundary.Compared with the blank control group,the cell morphology of ursolic acid group changed gradually,with fewer cells,scattered distribution,sparse arrangement among cells,and some cells became round or oval.After the addition of autophagy inhibitors,ursolic acid combined with 3-m A further reduced the number of cells,sparsely arranged between cells,and significantly changed the morphology of cells.3.Compared with the blank control group,ursolic acid induced autophagy in A549 cells,induced the increase of autophagy-related genes,protein LC3 and Beclin-1 expression;decreased P62 expression(P <0.01);Compared with ursolic acid group,3-MA made autophagy-related genes,protein LC3,Beclin-1 expression Significantly decreased;P62 expression increased significantly(P<0.01).4.Compared with the blank control group,ursolic acid can induce the autophagy of A549 cells,increase the expression of LC3 ? / I and beclin-1,decrease the expression of P62(P < 0.01),decrease the expression of LC3 ? / I and beclin-1,and increase the expression of P62(P < 0.01).Conclusion: Ursolic acid can inhibit the proliferation of A549 cells,and its mechanism may be related to the regulation of autophagy related LC3 ? / I,beclin-1 and P62 genes and protein expression to induce autophagy in tumor cells.