Study On The Mechanism Of Action Of Jianpi Xiaoai Prescription Against Liver Metastasis Of Colon Cancer Based On Metastasis-related Factors In Tumor Microenvironmen

Objective:Study on the mechanism of Jianpi Xiaoai Prescription against liver metastasis of colon cancer based on metastasis related factors in tumor microenvironment.Methods:（1）The logarithmic growth HCT116 cells were randomly divided into Jianpi Xiaoai Prescription group and blank control group.To observe the effect of Jianpi Xiaoai Prescription on the concentration of secreted exosomes of HCT116 cells and the expression of mi R-17 and mi R-21 in exosomes.（2）To observe the effects of Jianpi Xiaoai Prescription on the proliferation,invasion and migration of HCT116 cells.And the effect on the expression of tumor microenvironment metastasis-related factors:CXCR4,TGF-β,S100A4,S100A8,S100A9,ITGa_vβ5 in HCT116 cells.（3）The liver metastasis model of nude mice with colon cancer was established by injecting HCT116 cells through the spleen.The effects of different doses of Jianpi Xiaoai Prescription on liver metastasis and the expression of CXCR4,TGF-β,S100A4,S100A8,S100A9 and integrin a_vβ5 in the liver tissue of the model were observed.Results:（1）In the identification of exosomes,the concentration of exosomes in the blank control group was 1.2×10^{^9}particles/ml and that in the Jianpi Xiaoai Prescription group was 5.9×10^⁸particles/ml.There was significant difference between the two groups（p<0.05）.Compared with the control group,Jianpi Xiaoai Prescription group could significantly inhibit the expression of mi R-17 and mi R-21 in exosomes（p<0.05）.（2）Compared with the blank control group,the proliferation,invasion and migration of HCT116 cells in Jianpi Xiaoai Prescription group were decreased（p<0.05）.Compared with the blank control group,the protein and m RNA expressions of tumor microenvironment metastasis related factors including CXCR4,TGF-β,S100A4,S100A8,S100A9 and integrin a_vβ5 in Jianpi Xiaoai Prescription group were decreased（p<0.05）.（3）The statistical results showed that the liver metastasis area of the high-dose group was less than that of the model group,and the difference was statistically significant（p<0.05）.Compared with the model group,the metastases in the medium-dose and low-dose groups were also less,but there was no statistical significance（p>0.05）.Compared with the model group,the protein expression of S100A4 in liver tissue of high dose group was significantly decreased（p<0.01）.Compared with the model group,the protein expression of S100A8 was significantly decreased in the high,medium and low dose groups（p<0.01）.Compared with model group,the expression of CXCR4,TGF-β,S100A9 and integrin a_vβ5 in nude rat liver metastasis model were decreased in high and medium dose groups（p<0.05）.Conclusion:（1）Jianpi Xiaoai Prescription can inhibit the proliferation,invasion and migration of colon cancer HCT116 cells,and its mechanism may be by reducing the concentration of exosomes in colon cancer HCT116cells,and at the same time reducing the expression of metastasis-related factors in the tumor microenvironment,these factors are exosomal mi R-17,exosomal mi R-21,CXCR4,TGF-β,ITGa_vβ5,S100A4,S100A8 and S100A9.（2）Jianpi Xiaoai Prescription can inhibit liver metastasis of colon cancer nude mice,and its mechanism may be to reduce the expression of metastasis-related factors in the tumor microenvironment,these factors are CXCR4,TGF-β,ITGa_vβ5,S100A4,S100A8 and S100A9.