Study On Jianpi Yangzheng Xiaozheng Recipe Inhibiting Gastric Cancer EMT Through TAM

Aims:We sought to observe the influence of Jianpi Yangzheng Xiaozheng decoction Recipe and its disassembled groups on tumor-associated macrophages (TAM) infiltration and differentiation phenotype in transplanted tumor of MFC on 615 mice. Meanwhile, we also aimed to explore Changes of EMT and Key Molecules in Gastric, based on the changes in TAM phenotype.Methods:1. We establish a 615 mouse model with transplanted gastric cancer. The model mice were divided into 5 groups:the control group,5-Fu group, Jianpi Yangzhengxiao Recipe (whole compound group), Jianpi Yangzheng group, Xiaozhengsanjie group. We observe the inhibitory effect of all groups on subcutaneous tumor suppression and lung metastasis of gastric cancer.2. We examined the effect of Jianpi Yangzhengxiao Recipe and its disassembled groups on the number and phenotype of tumor-associated macrophages (TAM) in mice by flow cytometry and immunohistochemistry.3. We detected the effect of Jianpi Yangzhengxiao Recipe and its disassembled groups on EMT and PI3K/AKT signaling pathway proteins by real-time PCR, immunohistochemistry and Western Blot.Results:1. Compared with the control group, the whole compound group and the Jianpi Xiaozheng group could inhibit the tumor growth of mice (P<0.05). In the lung metastasis inhibition rate, the 5-Fu group> the whole compound group> the Xiaozao Sanjie group> the Jianpi Yangzheng group> the control group.2. Flow cytometry showed that:compared with the control group, Jianpi Yangzheng group can increase the expression of Ml type TAM (P<0.05), other groups can increase the number of Ml type TAM, but there was no statistical difference. Compared with the control group, all the groups can decrease the expression of M2 type TAM (P O.05).Immunohistochemistry showed that, the number of TAM (CD68+) was decreased in the whole compound group compared with the control group (P<0.05), the disassembled groups can decrease the number of TAM, but there was no statistical difference. Compared with the control group, the expression of M2 type TAM (CD 163+) of the Jianpi Yangzheng group was decreased (P<0.05), and the expression of CD 163 decreased in the other groups, but there was no statistical difference. Compared with the control group, the expression of M2 type TAM (CD206+) in all groups were decreased (P<0.05).3. Real-time quantitative PCR, immunohistochemistry and Western Blot showed that:Compared with the control group, whole compound group and its disassembled groups down-regulated N-cadherin, twist, Slug, Snail, mmp2,mmp9 and up-regulated E-cadherin.The effect of the whole compound group was better than that of the disassembled groups.The results of Western Blot showed that:compared with the control group, the expression of P-AKT, P-PI3K and P-MTOR were down-regulated in the all groups. The effect of the whole compound group was better than that of the disassembled groups.Conclusions:The effect of the whole compound group and the Jianpi Xiaozheng group can inhibit the growth of gastric cancer and the lung metastasis of the 615 mice. The whole compound group can inhibit the quantity of TAM. Jianpi Yangzheng group can increase the expression of M1 type TAM. The whole compound group and the disassembled groups can decrease the expression of M2 type TAM, and the effect of Jianpi Yangzheng Recipe is better than the whole compound group. The whole compound group and the disassembled groups can inhibit gastric EMT transformation and PI3K/AKT signaling pathway activation, and the effect of the whole compound is better than the disassembled groups.The whole compound group can ultimately reduce gastric cancer growth and metastasis, by reducing the number of TAM and M2 phenotype, gastric cancer EMT transformation...