Biochanin A Inhibits Glioblastoma Growth Via Restricting Glycolysis And Tochondrial Oxidative Phosphorylation

Object: To study the anticancer effects and the energy metabolism regulation of Biochanin A（BCA）in GBM,and explore the molecular mechanism of these effects.Methods: 1.U251 and U87 cells were selected as study objectives,and treated by different dosage of BCA.The activity of GBM cells was detected by CCK-8.Cellular proliferation was measured via an Edu assay and Colony formation.Apoptosis was detected by flow cytometry and the expression of apoptosis related proteins were evaluated by WB.In vivo intervention by use of BCA,pathological changes was observed by HE staining.Immunohistochemistry was used to observe the expression of Ki-67 protein.2.The ROS level were observed with a fluorescence microscope by DCF-DA staining.Mitochondria morphology was observed by mito-Tracker staining and transmission electron microscope.Western blotting analysed the relative proteins of mitochondrial fusion division.3.Mitochondrial stress test to detect mitochondrial energy metabolism and respiratory functions.Glycolytic stress test to detect glycolytic activities.Western blotting analysed the relative proteins of Glycolytic and Mitochondrial respiratory chain.Results: 1.BCA inhibited the proliferation of GBM cells in a time-and dosedependent manner.BCA increased the percentages of apoptosis and decreased apoptotic protein Bcl-2/Bax ratio.Meanwhile,HE staining showed looser tumor tissue of mice with BCA treatment.The expression of Ki-67 was significantly decreased in the BCA treatment group.2.The ROS level was increased significantly in BCA treatment groups.BCA increased mitochondrial fission.It promoted the mitochondrial division proteins Drp1 and decreased mitochondrial fusion proteins MFN1 and MFN2.3.BCA decreased the Mitochondrial Oxidative Phosphorylation and the Glycolytic Capacity of U251 Cells.It decreased the expression of mitochondrial respiratory chainrelated proteins,including ND1,SDHB,ATP5 A,and Glycolytic-related proteins,including p-AKT,p-m TOR,HIF-1α,GLUT1,HK2 and LDHA.Conclusions: BCA can inhibit the proliferation and induce apoptosis of glioblastoma cells.BCA can restrict glycolysis and mitochondrial oxidative phosphorylation in glioblastoma.