Study On The Efficacy And Mechanism Of Xianlian Jiedu Decoction In Inhibiting Liver Metastasis Of Colorectal Cancer Based On Tumor Microenvironment

OBJECTIVE:To observe the effect of Xianlian Jiedu Recipe on the proliferation and migration of colorectal cancer SW480 cells,and to explore the intervention effect of Xianlian Jiedu Recipe on liver metastasis of colorectal cancer in CT26 tumor bearing mice from the perspective of tumor microenvironment.Objective to provide scientific basis for the clinical application of Xianlian Jiedu Decoction in the prevention and treatment of colorectal cancer liver metastasis.Method:1.Effect of Xianlian Jiedu Recipe on proliferation and migration of colorectal cancer cells SW480 cells were stimulated with different concentrations of Xianlian Jiedu recipe.The effect of Xianlian Jiedu Recipe on SW480 cell morphology was observed under inverted microscope;MTT assay was used to detect the inhibitory effect of Xianlian Jiedu formula on the proliferation of colorectal cancer cells;DAPI staining was used to observe the effect of Xianlian Jiedu formula on the nuclear morphology of colorectal cancer;The effect of Xianlian Jiedu Recipe on apoptosis of colorectal cancer cells was observed by flow cytometry;Cell scratch test and Transwell test were used to observe the effect of Xianlian Jiedu formula on the migration of colorectal cancer cells.2.Study on the mechanism of Xianlian Jiedu Decoction on the migration of colorectal cancer cellsThe expression levels of E-cadherin,N-cadherin,vimentin,twist and snail in EMT of colorectal cancer cells were detected 48 hours after Xianlian Jiedu Decoction intervention to study the effect of Xianlian Jiedu Decoction on EMT of colorectal cancer cells.At the same time,analyze the protein expression levels of β-catenin,GSK-3β,P-GSK-3β,and CD44 in the Wnt/β-catenin signaling pathway,and explore whether Xianlian Jiedu Decoction can inhibit the Wnt/β-catenin signaling pathway and thereby inhibit the outcome.Metastasis of rectal cancer.3.Study on the efficacy and mechanism of Xianlian Jiedu Recipe on liver metastasis of colorectal cancer in miceConstruct a CT26 tumor-bearing mouse colorectal cancer liver metastasis model,intervene with different concentrations of Xianlian Jiedu Decoction,record the weight change of the mice during the administration period,and observe the number of tumor nodules and organ index in the liver of the mice after the mice are killed,Histopathological changes(HE staining),immunohistochemical method to detect TGF-β and CD44 expression in liver tissue,Western Blot method to detect FOXP3 expression in spleen tissue,flow cytometry to detect CD4+in mouse spleen The expression of CD25 and FOXP3 in T cells.Results:1.Through the MTT experiment and cell morphology changes,we found that Xianlian Jiedu Decoction has a significant inhibitory effect on the proliferation of colorectal cancer cells;2.DAPI staining and flow cytometry can show that Xianlian Jiedu Decoction can change Cell morphology and induce apoptosis of colorectal cancer cells;3.Scratch and Transwell experiments show that Xianlian Jiedu Decoction can inhibit the migration of colorectal cancer cells;4.Western Blot confirms that Xianlian Jiedu Decoction can inhibit colorectal cancer.EMT,its mechanism of action may be related to Wnt/β-catenin;5.Through mouse body weight,tumor nodule number,histopathology,TGF-β,CD44 changes in liver tissue,FOXP3 expression and apoptosis,it can be It is concluded that Xianlian Jiedu Recipe has an inhibitory effect on the liver metastasis model of colorectal cancer in mice,and its mechanism of action may be related to Treg cells.Conclusion:In vitro experiments show that Xianlian Jiedu Recipe can inhibit the proliferation and migration of colorectal cancer cells through the tumor microenvironment,and its mechanism may be inhibiting the occurrence of EMT by inhibiting the Wnt/β-catenin signaling pathway;mouse model experiments show that:Xian Lianjiedu Decoction has the effect of inhibiting liver metastasis of colorectal cancer,and its mechanism may be related to inhibiting the expression of Treg cells.