Study On The Mechanism Of Dapagliflozin Inhibites The Proliferation Of Breast Cancer Cells

ObjectiveTo explore the mechanism of the new hypoglycemic drugdapagliflozin in inhibiting the proliferation of breast cancer cells,so asto provide more theoretical basis for the treatment of breast cancerpatients with diabetes mellitus.Methods1.The expression of SGLT2 in human breast cancer and its adjacent tissues was observed by TCGA database analysis and immunohistochemistry.2.The gene microarray technology was used to compare and observe the changes of MCF-7 gene expression in human breast cancer cells after dapagliflozin treatment,and then using qRT-PCR and Western Blot(WB)to screen the possible targets of dapagliflozin.3.Flow cytometry was applied to assess the effect of dapagliflozin on the cell cycle of MCF-7 and MDA-MB-231 cells;WcB was performed to detect changes in the expression level of cell cycle-related proteins and P38-Mitogen Activated Protein Kinase(P38-MAPK)signal pathway-related proteins after dapagliflozin treatment.4.Using cell transfection technology to down-regulated or overexpressed the ERCC6 L in MCF-7 and MDA-MB-231 cells,and WB was used to detect the expression level of cell cycle-related proteins and P38-MAPK signaling pathway-related proteins.Results1.The TCGA database analysis and immunohistochemical staining observations showed that there was no significant difference in the SGLT2 expression between human breast cancer tissues and adjacent tissues.2.The results of gene microarray technology showed that after treatment with dapagliflozin in MCF-7 cells,the differentially expressed genes were mainly enriched in cell cycle related events;The qRT-PCR and WB results verified that dapagliflozin significantly inhibited the expression of ERCC6 L in MCF-7 and MDA-MB-231 cells.3.The flow cytometry and WB results showed that after treating breast cancer cells with dapagliflozin,the number of cells in G1 phase increased,while that in S phase and G2 phase decreased,and the expression of cyclin D1,cyclin E1,p-p38 and Fos were decreased,while the expression of DUSP1 were increased.4.After down-regulating the expression of ERCC6 L in breast cancer cells,the expression of Cyclin D1,Cyclin E1,p-P38 and FOS was down-regulated,and the expression of DUSP1 was increased.After overexpression of ERCC6 L in breast cancer cells,compared with the NC group,the ERCC6 L expression was up-regulated in the ERCC6 L overexpression group,while the the ERCC6 L expression was significantly decreased in ERCC6 L and dapagliflozin co-treatment groups.ConclusionDapagliflozin can block the DUSP1-P38-MAPK-FOS signaling pathway by down-regulating the expression of ERCC6 L,thereby inhibiting the cell cycle and proliferation of breast cancer cells.