Baicalin Inhibits The Invasion And Epithelial-mesenchymal Transition Of Triple-negative Breast Cancer And Its Related Mechanism

Part 1.Effect and Mechanism of Baicalin on Migration,Invasion and Epithelial-mesenchymal Transition of Triple-negative Breast Cancer MDA-MB-231 CellsObjective: To observe the effect of LPS on the proliferation,migration,invasion and epithelial-mesenchymal transition of breast cancer MDA-MB-231 cells,and explore the inhibitory effect of baicalin on LPS-induced breast cancer MDA-MB-231 cells.Methods: Different concentrations(0,5,10,20,40?g/mL)of LPS added to breast cancer MDA-MB-231 cells for 24 h,48h,72 h,The effects of LPS on the proliferation,migration,invasion and markers of epithelial-mesenchymal transition in breast cancer MDA-MB-231 cells were detected by CCK8,scratch test,Transwell chamber assay,immunofluorescence assay,Western blot and q PCR.Induction of breast cancer MDA-MB-231 cells by LPS,detection of baicalin on proliferation,migration,invasion and epithelial to mesenchymal transition of breast cancer MDA-MB-231 cells by CCK8,Scratch test,Transwell chamber assay,Western blot and q PCR The impact of things.Results: 1.After different concentrations of LPS were applied to breast cancer MDA-MB-231 cells,the morphology of cells changed from the phenotype of epithelial cells to the phenotype of mesenchymal cells.2.LPS can promote the proliferation of breast cancer MDA-MB-231 cells with a concentration and time-dependent,48 h,20?g/m L is the best time and concentration,cell proliferation rate of 84.8% ± 3.11%(P<0.05).3.LPS could enhance the migration of breast cancer MDA-MB-231 cells with the concentration and time-dependent,48 h,20?g/m L was the best time and concentration,and the cell migration rate was 91.47%±3.10%(P<0.05).4.LPS could increase the invasion of breast cancer MDA-MB-231 cells,and the number of cell invasion in 20 ?g/m L group was 158.67±9.07(P<0.05).5.Immunofluorescence experiments suggest that the expression of ?-catenin is mainly located in the nucleus.6.After treatment with LPS,the expression of Vimentin and ?-catenin protein and m RNA in breast cancer MDA-MB-231 cells were up-regulated(P<0.05).The expression of E-cadherin protein and m RNA was down-regulated(P<0.05).And correlated with the concentration.LPS 20 ?g/m L The group was the most significant(P<0.05).7.CCK8 assay showed that 72 h,40?g/m L baicalin group inhibited the proliferation of breast cancer MDA-MB-231 cells,the inhibition rate was 79.2% ± 3.11%(P <0.05).8.Compared with the model control group and baicalin different concentration group,40?g/m L baicalin group inhibited the migration rate of breast cancer MDA-MB-231 cells was 37.10% ± 3.60%(P <0.05),the number of cell invasion was 45.67 ± 5.13(P<0.05).9.Compared with the model control group and baicalin different concentration group,the relative expression of E-cadherin protein in breast cancer MDA-MB-231 cells in the 40?g/m L baicalin group was 0.702±0.052(P<0.05),and the relative expression of Vimentin protein was 0.798± At 0.025(P<0.05),the relative expression of ?-catenin protein was 0.752±0.038(P<0.05),the relative expression of MMP-7 protein was 0.752±0.056(P<0.05),and the relative expression of c-Myc protein was 0.765±0.077(P<0.05),the relative expression of E-cadherin m RNA was 0.787±0.046(P<0.05),the relative expression of Vimentin m RNA was 0.742±0.107(P<0.05),and the relative expression of ?-catenin m RNA was 0.794±0.071(P<0.05).The relative expression of MMP-7 m RNA was 1.739±0.028(P<0.05),and the relative expression of c-Myc m RNA was 1.731±0.073(P<0.05).Conclusion: LPS can enhance the proliferation,migration and invasion of breast cancer MDA-MB-231 cells in vitro;LPS can induce epithelial-mesenchymal transition in breast cancer MDA-MB-231 cells in vitro;LPS effect on breast cancer MDA-MB-231 cells It may play a role through the Wnt/?-catenin signaling pathway.Baicalin can inhibit the proliferation,migration and invasion of breast cancer MDA-MB-231 cells in vitro,and up-regulate the expression of E-cadherin and down-regulate the expression of Vimentin,?-catenin,c-Myc and MMP-7.Part 2.The Effect and Mechanism of Baicalin on Three-negative Breast Cancer Transplantation Model RatsObjective: To investigate the effect of baicalin against triple negative breast cancer in vivo and its effect on downstream target molecules of Wnt/?-catenin signaling pathway.Methods: In the in vivo experiment,a tumor-bearing mouse model of breast cancer was prepared,and the general condition,weight,and weight of the transplanted tumor were observed.The pathological changes were detected by the HE method.E-cadherin,Vimentin was detected by immunohistochemistry,Western blot,and q PCR.Expression of ?-catenin,c-Myc,and MMP-7.Results: 1.In vivo experiments,successfully prepared nude mice breast cancer xenograft model,baicalin high-dose group(200mg/kg)can improve the general state of tumor-bearing mice,reduce the weight of xenografts(P<0.05).2.Compared with the model control group and baicalin different dose group,the average integrated optical density of E-cadherin in the high dose group of baicalin was 70.57±4.75(P<0.05),and the average integrated optical density of Vimentin was 56.57±3.77(P<0.05),the average integrated optical density of ?-catenin was 59.45±5.44(P<0.05),the average integrated optical density of MMP-7 was 44.11±5.13(P<0.05),and the average integrated optical density of c-Myc was 40.47±2.72(P< 0.05).<0.05).3.Compared with the model control group and baicalin different dose group,the relative expression of E-cadherin m RNA in the high dose group of baicalin was 0.789±0.062(P<0.05),and the relative expression of Vimentin m RNA was 0.839±0.075(P<0.05).The relative expression of ?-catenin m RNA was 0.915±0.064(P<0.05),the relative expression of MMP-7 m RNA was 1.834±0.073(P<0.05),and the relative expression of c-Myc m RNA was 1.563±0.128(P<0.05).4.Compared with the model control group and the baicalin different dose group,the relative expression of E-cadherin in the high dose group of baicalin was 0.684±0.033(P<0.05),and the relative expression of Vimentin was 0.824±0.079(P<0.05).The relative expression of ?-catenin protein was 0.698±0.058(P<0.05),the relative expression of MMP-7 protein was 0.768±0.109(P<0.05),and the relative expression of c-Myc protein was 0.794±0.068(P<0.05).Conclusion: Baicalin can inhibit the growth of triple-negative breast cancer xenografts in vivo,up-regulate the expression of E-cadherin,down-regulate the expression of Vimentin,?-catenin,c-Myc,MMP-7,and regulate the Wnt/?-catenin signal.The target is related to downstream pathways.