Oroxylin A Inhibits The Proliferation Of Triple-negative Breast Cancer Cell Line MDA-MB-231

[Objective]To study the effect and mechanism of oroxylin A on proliferation of triple-negative breast cancer cell line MDA-MB-231.[Methods](1)Network pharmacology was used to predict the targeted genes,functional enrichment and pathway enrichment of oroxylin A in triple-negative breast cancer.(2)Incubating MDA-MB-231 cell with oroxylin A in 0,200?M for24 h,optical microscope was carried out to observe the number and morphology.(3)MDA-MB-231 cells were treated with oroxylin A of 0,50,100,150 and 200?M for 24 h,48h and 72 h,CCK-8 was carried out to detect the proliferation inhibition rate of the drug.(4)After treatment of MDA-MB-231 cells with 0,100,150,200?M oroxylin A for 24 h,and after 24,48,and 72 hours of treatment with 150?M,Annexin V-FITC and PI double staining were used for flow cytometry analysis.(5)Genes and proteins expression levels of BAX,BCL2 and BECN1 proteins were detected by RT-PCR and Western blotting.[Results](1)Network pharmacology analysis showed that oroxylin A has 12 functional targets in triple-negative breast cancer.The results of enrichment analysis showed that oroxylin A acted on multiple signaling pathways of cells to perform a variety of biological functions.(2)The results of microscopic observation showed that the number of living cells was less and the morphology of cells changed after treatment with oroxylin A.(3)CCK-8 results showed that cell proliferation was significantly inhibited in the concentration group compared with the control group(P<0.01).48 h group compared with control group,beginning from 100?M,cell proliferation was significantly inhibited(P<0.05).72 h group compared with control group,beginning from 50?M,cell proliferation was significantly inhibited(P<0.05).The half inhibition rate at 24 h was(146.66±7.59)?M.(4)The results of flow cytometry showed that oroxylin A promoted cell apoptosis.Compared with control group,the proportion of apoptotic cells in all other groups increased significantly(P<0.05).(5)RT-PCR and Western blotting results showed that oroxylin A promoted the expression of BAX and BECN1,inhibited the expression of BCL2,and up-regulated the level of BAX/BCL2(P<0.05).[Conclusions]Oroxylin A induced apoptosis of MDA-MB-231 cells by up-regulating BAX expression,down-regulating BCL2 expression,increasing the ratio of BAX to BCL2.Oroxylin A promoted autophagy of MDA-MB-231 cells by up-regulating BECN1 expression.The effect of oroxylin A is concentration dependent and time dependent.