The Role Of Aerobic Glycolysis In Histone Deacetylase Inhibitor Induced Breast Cancer Cell Death

BackgroundBreast cancer is one of the most common malignant tumors in women all over the world.The onset age of breast cancer is younger,which seriously endangers women’s health.There are many types of breast cancer,among which the treatment of triple negative breast cancer is the most difficult.Therefore,finding new therapeutic targets and combining drugs are the key to improve the survival time of triple negative breast cancer.Studies have shown that histone deacetylase plays an important role in tumorigenesis and development,but the relationship between histone deacetylase and breast cancer has been less reported,and its possible regulatory mechanism is still unclear.ObjectiveTo clarify the effect of histone deacetylase inhibitors on the survival and apoptosis of breast cancer cells and to explore the role of aerobic glycolysis in cytotoxicity induced by histone deacetylase inhibitors,so as to provide new molecular targets and combined drug strategies for clinical treatment of breast cancer.MethodsTri-negative breast cancer cells(BT-549 and MDA-MB-231)were cultured in vitro and treated with histone deacetylase inhibitor LBH589.CCK8 assay was used to detect the effect of LBH589 on the survival rate of breast cancer cells.Flow cytometry was used to detect the change of apoptotic rate.Western blotting and cleaved caspase-3 expression levels were detected.The level of aerobic glycolysis was detected by glucose uptake and lactic acid production detection kit.Combined with 2-DG and 3-BP,CCK8 was used to detect the effect of combined action on cell viability.Western blotting was used to detect the expression level of Caspase-3 and flow cytometry was used to detect the change of apoptotic rate.Results1.CCK8 results showed that different concentrations of LBH589 could significantly reduce the survival rate of breast cancer cells(BT-549 and MDA-MB-231),with significant difference(P< 0.05).2.Flow cytometry showed that different concentrations of LBH589 could significantly increase the apoptotic rates of BT-549 and MDA-MB-231 cells.3.Western blotting results showed that different concentrations of LBH589 could significantly increase cleaved caspase-3 expression in breast cancer cells BT-549 and MDA-MB-231(P< 0.05).4.The results of wine uptake and lactic acid production test kit showed that compared with the control group,the glucose uptake and lactic acid production of BT-549 and MDA-MB-231 cells were significantly increased by LBH589 without concentration(p < 0.05).5.CCK8 results showed that the combination of 2-DG and 3-Bromopyruvate(3-BP)could further increase the cell viability of BT-549 and MDA-MB-231 cells induced by LBH589(P< 0.05).6.Western blotting results showed that the combination of 2-DG and 3-BP could further increase the expression of cleaved caspase-3 induced by LBH589(p < 0.05)and MDA-MB-231(P< 0.05).7.Flow cytometry showed that the combination of 2-DG and 3-BP could further increase the apoptotic rates of BT-549 and MDA-MB-231 induced by LBH589.ConclusionHistone deacetylase inhibitor LBH589 can significantly reduce the survival rate and increase apoptosis of breast cancer cells,and combined with aerobic glycolysis inhibitor can further increase the cytotoxicity of LBH589.The study provides a new strategy for the clinical treatment of breast cancer.