Exploring the mechanism of icariin regulating ferroptosis in colon cancer cells based on the MDM2/P53/SLC7A11 signaling pathway

To investigate the mechanism of action of icariin in regulating ferroptosis in colon cancer cells via the mouse double minute 2 (MDM2)/tumor protein 53 (P53)/solute carrier family 7, member 11 (SLC7A11) signaling pathway.

The core target of iicariin to promote ferroptosis in colon cancer cells was predicted and verified using network pharmacology and molecular docking. The experiment included the vehicle control group, icariin low-concentration group (5 μmol/L), icariin medium-concentration group (15 μmol/L), and icariin high-concentration group (25 μmol/L), ferroptosis promoter group, empty vector control group, MDM2 overexpression group, and MDM2 overexpression + icariin medium-concentration group. The MDM2 overexpression and control plasmids were transfected into human colon cancer HCT116 cells. Cell proliferation was assessed using the CCK-8 assay. The concentration of Fe²⁺, glutathione (GSH), and malonaldehyde (MDA) was detected using cobrimetric method, microplate method and TBA method. The expression level of reactive oxygen species (ROS) was detected using immunofluorescence. The expression level of MDM2, P53, SLC7A11, PTGS2, and GPX4 protein and mRNA was detected using western blotting and RT-qPCR, respectively.

The 12 targets of icariin affecting ferroptosis in colon cancer were identified by network pharmacology, among which 4 core targets were MDM2, EGFR, PARP1, and SRC. Icariin has a good binding activity with MDM2 was revealed by molecular docking.Icariin regulates ferroptosis in colon cancer cells through MDM2. Compared with the vehicle control group, the proliferative activity, GSH concentration, MDM2, SLC7A11, and GPX4 protein and mRNA expression of cells decreased in the icariin medium-concentration and high-concentration groups (P < 0.01); MDA and Fe²⁺ concentrations, ROS levels, and P53 and PTGS2 protein and mRNA expression increased in the icariin medium-concentration and high-concentration groups (P < 0.05, P < 0.01). Compared with the MDM2 overexpression group, cell proliferative activity, GSH concentration, MDM2, SLC7A11, and GPX4 protein and mRNA expression decreased in the MDM2 overexpression+icariin medium-concentration group (P < 0.01); MDA and Fe²⁺ concentrations, ROS levels, and P53 and PTGS2 protein and mRNA expression increased in the MDM2 overexpression+icariin medium-concentration group (P < 0.05, P < 0.01).

Icariin can promote ferroptosis in human colon cancer HCT116 cells, and its mechanism may involve inhibiting MDM2 expression and regulating the MDM2/P53/SLC7A11 signaling pathway.