Effect Of Realgar On Cell Proliferation,Invasion,Migration,and Cell Epithelial Stromal Transformation Of Esophageal Cancer Cells

OBJECTIVE: To observe the effect of realgar on the proliferation,migration and invasion of esophageal carcinoma cell line Eca109 and explore its mechanism.Methods: Different concentrations of realgar(0,10,20,40,60,80,100μmol · L-1)or realgar 1/2 IC50,IC50,2 IC50 were applied to esophageal cancer cells Eca109.The ability of different concentration gradients of realgar on the proliferation and colony formation of esophageal cancer cells Eca109 was observed by CCK-8 cell proliferation assay and cell colony formation assay,and the half maximal inhibitory concentration(IC50)of realgar on esophageal cancer cells was calculated for subsequent experimental grouping;the effects of different concentrations of realgar on the migration and invasion of esophageal cancer cells were observed by cell scratch assay and transwell chamber invasion assay;the effects of different concentrations on the expression of proteins related to tumor epithelial-mesenchymal transition of esophageal cancer cells were observed by Western blot assay and immunofluorescence assay to further explore the mechanism of realgar.RESULTS:(1)CCK-8 cell proliferation assay showed that realgar could effectively inhibit the proliferation of esophageal cancer cells in a time-and concentration-dependent manner(P<0.01).(2)Cell colony formation assay showed that the proliferation of Eca109 cells showed a decreasing trend dependent on the increase of realgar concentration,and the lower the colony formation rate of esophageal cancer cells(P<0.01);(3)Cell scratch assay showed that realgar could inhibit the migration ability of esophageal cancer cells in a concentration-dependent manner(P<0.01);(4)Transwell chamber invasion assay showed that realgar could reduce the number of cell invasion in a concentration-dependent manner(P<0.01);(4)Western blot assay showed that the relative expression levels of N-cadherin and Slug protein in esophageal cancer cells of realgar 1/2 IC50 group,IC50 group and2IC50 group were lower than those of blank group(P<0.01),and the relative expression levels of E-cadherin protein were higher than those of blank group(P<0.01);(3)Immunofluorescence staining results showed that compared with blank group,the expression levels of N-cadherin and Slug in Eca109 cells of realgar IC50group and 2IC50 group were down-regulated,and the expression levels of E-cadherin were up-regulated(P<0.01);And compared with blank group,the expression levels of N-cadherin and Slug in Eca109 cells of realgar 1/2IC50 group were down-regulated(P<0.05),and the expression levels of E-cadherin were no changes(P>0.05)CONCLUSION: Realgar can effectively inhibit the proliferation,migration and invasion of esophageal carcinoma cell line Eca109,and also inhibit the epithelial-mesenchymal transition of tumor cells.