Study On Mechanism Of Yanghe Decoction In Inhibiting Human Breast Cancer Cell Migration And Invasion Based On C-Met/NF-κB/CXCL1 Signaling Pathway

Objective: To study the molecular biological mechanism of Yanghe Decoction in the intervention of human breast cancer cell migration and invasion in vitro,to further clarify the regulatory role of c-Met/NF-κB/CXCL1 in breast cancer,to provide new ideas for the molecular targeting treatment of breast cancer in the future,and to explore the clinical application value of Yang he Decoction in the treatment of breast cancer.Methods:(1)After the treatment of MDA-MB-231 cell 24 h with different concentrations of Yanghe Decoction containing serum,Transwell cell invasion experiment and scratch healing experiment were used to study the effect of Yanghe Decoction serum on the ability of MDA-MB-231 invasion,migration and repair of breast cancer cells.Western blot was used to detect the change of the expression of total c-Met and p-c-Met and NF-κB p65 protein in MDA-MB-231 cells.The expression of CXCL1 was detected by ELISA.(2)c-Met inhibitor SGX523 was used to reduce the expression of c-Met,and to detect the effect of Yanghe Decoction serum on the effect of SGX523,and then to study whether the influence of Yanghe Decoction containing serum on MDA-MB-231 was realized by regulating c-Met.Transwell cell invasion experiment and scratch healing test were used to detect the influence of Yanghe Decoction containing serum on the invasion,migration and repair ability of MDA-MB-231 cells after SGX523 application.Western blot was used to detect the expression of total c-Met,p-c-Met,NF-κB p65 protein in the cells.ELISA assay was used to detect the expression of CXCL1.(3)Using the NF-κB inhibitor PDTC,the expression of NF-κB was downregulated,and the effect of Yanghe Decoction serum on the effect of PDTC was detected,and the effect of Yang He Decoction serum on MDA-MB-231 was studied by regulating NF-κB.Transwell cell invasion experiment and scratch healing test were used to detect the influence of Yanghe Decoction containing serum on the invasion,migration and repair ability of MDA-MB-231 cells after PDTC application.Western blot was used to detect the expression of total c-Met,p-c-Met,NF-κB p65 protein in the cells.ELISA assay was used to detect the expression of CXCL1.Results:(1)With the increase of serum concentration of Yanghe Decoction,the effect of serum inhibiting cell invasion and migration of Yanghe Decoction increased gradually.At the same time,the expression of total c-Met,p-c-Met,NF-κB p65 and CXCL1 protein in MDA-MB-231 cells decreased significantly,showing a concentration dependence.(2)The use of c-Met inhibitor SGX523 can enhance the inhibitory effect of Yanghe Decoction containing serum on cell invasion and migration,and enhance the down-regulation of total c-Met,p-c-Met,NF-κB p65 and CXCL1 protein expression in the cells.(3)The use of NF-κB inhibitor PDTC can enhance the inhibitory effect of Yanghe Decoction containing serum on cell invasion and migration,and enhance the down regulation of NF-κB p65 and CXCL1 protein expression in the cells,but there is no obvious effect on the expression of total c-Met and p-c-Met,suggesting that c-Met is the upstream of NF-κB p65.Conclusions:(1)Yanghe Decoction containing serum can inhibit the activation of c-Met and NF-κB in human breast cancer cell MDA-MB-231 and down regulate the protein expression level of c-Met,NF-κB and its downstream CXCL1;(2)The mechanism of Yanghe Decoction inhibiting the migration and invasion of human breast cancer cell MDA-MB-231 may be achieved by inhibiting the activity of c-Met/ NF-κB /CXCL1.