Curdione regulates ferroptosis in colorectal cancer via N6-methyladenosine

Abstract

Abstract Background: Curdione is a sesquiterpene isolated from Curcumae Rhizoma with high biological activity and a wide range of pharmacological effects. The traditional Chinese medicine Curcumae Rhizoma inhibits the development of many types of cancer, especially colorectal cancer, but the anti-colorectal mechanism of its monomer Curdione is vacant. Methods: CT26 cells were treated with 12.5 μM, 25 μM, and 50 μM of Curdione, and cell activity was measured by MTT. Nude mice were implanted subcutaneously with different doses of Curdione and oxaliplatin by tail vein injection, and tumor histology was examined by HE staining. Flow cytometry was used to detect ROS in cells and tissues. Kits were used to detect levels of iron ions, MDA and GSH. PCR and western blot were used to detect ferroptosis and m6A modification-related factors. The methylation spot hybridization assay detected quantitative changes in overall methylation. MeRIP-qRNA measured SLC7A11 and HOXA13. shRNA-METTL14 plasmid was constructed to verify the inhibitory effect of Curdione on colorectal cancer. Results: A dose-dependent decrease in activity was observed in Curdione-treated cells. Curdione increased the accumulation of ROS in colorectal cancer cells and tumor tissues, significantly increased the levels of MDA and Fe2+, and decreased the activity of GSH. qPCR and Western blot results showed that Curdione promoted the expression of METTL14 and YTHDF2 in colorectal cancer cells and tissues, and decreased the expression of SLC7A11, SLC3A2, HOXA13, and GPX4 expression. In addition, in animal experiments, HE staining showed that the Curdione -treated group showed significant necrosis of tumor cells. Moreover, the levels of m6A modifying factors, namely SLC7A11 and HOXA13, were increased in the tissues after the drug intervention compared to the control group. The knockdown of METTL14 was followed by an increase in CT26 cell activity and a decrease in reactive oxygen species. Cellular activity and GSH levels decreased after Curdione treatment. Levels of ROS, MDA, and iron ions increased significantly. Conclusion: These results suggest that Curdione induces ferroptosis in colorectal cancer via m6A methylation.