Flavonoid Glycoside Diosmin Induces Apoptosis and Cell Cycle Arrest in DLD-1 Human Colon Cancer Cell Line

Abstract

Abstract The purpose of this study was to see if diosmin induced apoptosis and inhibited cell proliferation in DLD-1 cell lines. The assays such as MTT, DCFDA, AO/EtBr, cell cycle analysis, annexin V-FITC apoptosis by flow cytometry, gene expression analysis were done to evaluate the in vitro anti-colon cancer potential of diosmin in DLD-1 human colon cancer cell line. While analyzing the results of MTT assay it is observed that when different concentrations of diosmin were induced, it was discovered that DLD-1 cells were killed in a dose-dependent manner. Diosmin mediated apoptosis induction is observed through AO/EtBr double staining method. Diosmin treatment in DLD-1 cell line generates high amount of ROS which is being analyzed using ROS DCFDA assay. While using flow cytometry to investigate the effect of diosmin on cell cycle distribution, it was found that diosmin-induced cell cycle arrest occurs in the S phase. When diosmin is administered, the proportion of surviving cells decreases, confirming the existence of cell death induction via apoptosis using the annexin V staining method. Diosmin upregulated mRNA levels expression of p53 and p21, indicating that diosmin induced S phase arrest and apoptosis in DLD-1 cells is controlled in part by the p21/p53 signaling pathway. GRAPHICAL ABSTRACT