The Mechanism Of Paris Polyphylla Based EMT On Colorectal Cancer Cell Migration And Invasion Is Discussed

Objective:In this study,Paris polyphylla and its monomers were selected to explore the mechanism of action of inhibiting colorectal cancer metastasis and invasion by regulating EMT,to clarify its inhibitory effect on colon cancer metastasis,so as to provide experimental support for drug targeting on the metastasis of colorectal cancer,and provides new ideas and methods for controlling the metastasis of colon cancer.Methods:（1）Culture human colorectal cancer HT29 and LoVo cell lines.（2）The inhibitory effect of different concentrations of extract of medication on the growth of HT29 and LoVo cells was detected by MTT assay,and the IC₅₀ value of medication was analyzed.Filter the components of medication,select the main monomers of medication for IC₅₀ value screening,and select the most sensitive monomers for follow-up experiments..（3）The effect of medication on cell proliferation was detected by plate cloning and Ed U proliferation assay;Western-blotting detected protein expression of the PI3K/Akt signaling pathway after medication intervention.（5）Transwell examined the effects of medication on cell migration and invasion;Western-blotting was used to detect the expression of EMT-related proteins after medication interventions.Results:（1）Human colorectal cancer HT29 and LoVo cells grew well and were suitable for experimental operation.（2）The results of MTT assay showed that PPE has an inhibitory effect on HT29 and LoVo cell growth in a concentration-dependent manner.（3）The results of plate cloning and Ed U proliferation experiments show that PPE has an inhibitory effect on cell proliferation.（4）Western-blotting detected the expression of PI3K/Akt-related proteins.The results showed that after PPE intervention in the cells,the expression levels of p-PI3K and p-Akt decreased compared with the control group.（5）Transwell results showed that PPE inhibited cell migration and invasion.（6）Western-blotting detected EMT-related proteins,and the results showed that after PPE intervention in cells,the expression levels of E-cadherin increased,the expression levels of N-cadherin,MMP2 and MMP9decreased,the expressions of Smad2 and Smad3 were basically unchanged,and the expression levels of p-Smad2 and p-Smad3decreased.（7）The contents of Polyphyllin I（PPI）,Polyphyllin II（PPII）,and Polyphyllin VII（PPVII）in the Paris polyphylla are the quality standard for detecting the Paris polyphylla,and the MTT results showed that PPI,PPII,PPVII had inhibitory effects on cell viability,PPVII has the most obvious inhibition on cell viability,so PPVII was selected for subsequent experiments.（8）The results of plate cloning and Ed U proliferation experiments showed that PPVII had an inhibitory effect on cell proliferation.（9）Western-blotting detected PI3K/Akt-related protein expression,and the results showed that after PPVII intervention in cells,the expression levels of p-PI3K and p-Akt decreased compared with the control group.（10）Transwell experiments showed that PPVII inhibits cell migration and invasion.（11）Western-blotting detected EMT-related proteins,and the results showed that after PPVII intervention in cells,the expression levels of E-cadherin increased,the expression levels of N-cadherin,MMP2 and MMP9decreased,the expressions of Smad2 and Smad3 were basically unchanged,and the expression levels of p-Smad2 and p-Smad3 decreased compared with the control group.