Study On The Effect Of Sanjie Xiaoliu Recipe On The Invasion And Migration Of Breast Cancer 4T1 Cells Based On TGF-β/Smads Pathwa

Objective: To explore the effect of Sanjie Xiaoliu recipe on breast cancer4T1 cells in vitro,and to explore the mechanism of Sanjie Xiaoliu recipe acting on TGF-β/Smads pathway and inhibiting epithelial interstitial transformation.Methods: Firstly,4T1 cells from breast cancer mice were cultured in vitro,and the cells in logarithmic growth phase were taken for related experiments.1.CCK-8 test was carried out and divided into 5%,10%,15%,20%,blank serum group and control group corresponding to different concentrations of drug-containing serum,to explore the effect of Sanjie Xiaoliu recipe drug-containing serum on the inhibition of 4T1 cell proliferation.2.According to the results of CCK-8 test,the intervention concentration of drug-containing serum of Sanjie Xiaoliu recipe on breast cancer 4T1 cells was determined,and then TRANSWELL migration test and TRANSWELL invasion test were carried out to compare the migration and invasion ability of4T1 cells after intervention with drug-containing serum.3.The 4T1 cells treated with different concentrations of Sanjie Xiaoliu recipe containing serum were collected.The phosphorylation of Smad2/Smad3 protein in TGF-β/Smads pathway and the levels of E-cadherin,N-cadherin and Vimentin related to epithelial-mesenchymal transformation in 4T1 cells were detected by Western blotting.Results:1.After the intervention of different concentrations of Sanjie Xiaoliu recipe containing serum on 4T1 cells by CCK-8 method,it was found that the 10% concentration of drug containing serum had the most stable inhibitory effect on 4T1 cells,and the inhibition rate gradually increased with time,while the 15% concentration of drug containing serum had the best effect on cell proliferation in 72 hours.The intervention concentration of drug-containing serum of Sanjie Xiaoliu prescription was 10% and 15%.2.TRANSWELL migration test showed that the movement ability of 4T1 cells in the 10% Sanjiexiaoliu prescription drug-containing serum group was significantly lower than that in the blank serum group.The number of cells in the Sanjie Xiaoliu prescription drug-containing serum group was 74.00 ±24.43,and the number of cells across the membrane in the blank serum group was172.30 ±37.87,and the migration rate was 42.94%.TRANSWELL invasion test showed that the invasive ability of 4T1 cells in the 10% Sanjiexiaoliu prescription drug-containing serum group was significantly lower than that in the blank serum group.The number of cells in the Sanjie Xiaoliu prescription drug-containing serum group was(86.80 ±64.01),and that in the blank serum group was(425.20 ±112.30),and the invasion rate was 20.41%(P < 0.001).3.Western blotting showed that the phosphorylation of Smad2/Smad3,the key protein in TGF-β/Smads signal pathway,was down-regulated in 10%,12.5%and 15% Sanjie Xiaoliu recipe containing serum intervention group(P < 0.05).The expression of E-cadherin N-cadherinand vimentin were up-regulated and down-regulated respectively.Conclusion:1.Sanjie Xiaoliu recipe can inhibit the proliferation of breast cancer 4T1 cells.2.Sanjie Xiaoliu recipe can inhibit the movement and invasion of breast cancer4T1 cells.3.The regulatory effect of Sanjie Xiaoliu recipe on EMT may be achieved by down-regulating the pathway TGF-β/Smads to reduce the level of interstitial phenotypic marker protein and increase epithelial phenotypic marker protein.