To Explore The Effect Of The Compatibility Of Astragalus And Curcuma On Tumor Cell Migration And Adhesion

Objective:In this study,the Chinese classic compatible drugs with qi-enriching Astragalus and blood-activating zedoary(a traditional Chinese medical Yiqi Huoxue method)is applied.And CT26,Lewis and Hepal-6 are choosed for in vitro experiments.After the treatment with the drugs,the protein expression and mRNA expression of the adhesion-related factors—E-cadherin,KAI1,PTEN,CD 147,HIF-1?,and MMP-9 in the three kinds of cells are detected,thus to investigate the effect of astragalus and zedoary on the migration and adhesion of tumor cells.Methods:1.The CCK-8 method was used to detect the effects of different concentrations of astragalus and zedoary on the activity of tumor cells,so as to select the appropriate dosage for follow-up experiments;2.Observe the healing effect of astragalus and zedoary on the treated tumor cells by scratch experiment to determine the effect of the drugs on tumor cell migration ability;3.By applying cell adhesion experiments,the effects of astragalus and zedoary on the adhesion between the treated tumor cells and intercellular adhesion and the adhesion to extracellular matrix are detected4.Western blot and Real-Time PCR are used to detect the protein expression and mRNA expression of adhesion-related factors—E-cadherin,KAI1,PTEN,CD147,HIF-1?,MMP-9 in tumor cells.Results:1.Astragalus and zedoary in each concentration group have different inhibitory effects on the activity of tumor cells.In order to minimize the effect of drugs on the activity of cells,the 0.0125,0.025 and 0.05g/ml concentration groups are selected for the follow-up experiment.2.The scratch experiment results show that the mobility of CT26 and hepal-6 cells significantly decrease after the drug treatment,and it is concentra-tiondependent.Th e mobility of Lewis cells is only significantly reduced in the 0.05 g/ml group.3.The results of cell adhesion experiments show that the adhesion rate between CT26 and Hepal-6 cells is significantly increased in a drug-dependent manner,and the concentration is dependent.Only 0.025 g/ml of Lewis cells show significant increase in intercellular adhesion;The adhesion rate of CT26,hepal-6 and Lewis cells to extracellular matrix decreases significantly after drug treatment,and shows a concentration dependence.4.After drug treatment,the expressions of protein and mRNA of e-cadherin,PTEN and KAI1 in CT26 and Lewis cells are up-regulated;and the expressions of protein and mRNA of CD 147,HIF-1 and MMP-9 are down-regulated;The protein and mRNA expressions of e-cadherin,PTEN and KAI1 in hepal-6 cells are up-regulated;the protein and mRNA expressions of CD147 are down-regulated;and the mRNA expressions of HIF-1 and MMP-9 are down-regulated.Conclusion:the compatibility of astragalus and zedoary can reduce the migration ability of CT26 and Hepal-6 cells,and there is uncertainty about the influence on the migration ability of Lewis cells.It can enhance the adhesion between CT26 and hepal-6 cells and their intercellular adhesion,and reduce the adhesion ability of CT26,Lewis,hepal-6 cells and extracellular matrix.As for the ability of Lewis cells and their intercellular adhesion,it remains to be further explored.The underlying mechanism affecting cell migration and adhesion ability may be related to the up-regulation of adhesion related factors E-cadherin,KAI1,PTEN and the down-regulation of CD147,HIF-1 and MMP-9.