Gingerenone A induces ferroptosis in colorectal cancer via targeting suppression of SLC7A11 signaling pathway

Background: Colorectal Cancer (CRC) is one of the most common and fatal diseases, yet effective therapeutic drugs are lacking in clinical settings. Gingerenone A (GA) is an active compound derived from ginger, has demonstrated anti-tumor properties. However, the efficacy of GA against CRC and its primary mechanism of action remain unclear.

Materials and methods: MTT assay and colony formation assay were employed to evaluate cell viability. Transwell assays were utilized to assess the migratory and invasive capabilities of the cells. The effects of GA on Ferroptosis related proteins were analyzed using Western blot. Levels of glutathione (GSH), malondialdehyde (MDA), Fe²⁺, and 4-hydroxynonenal (4-HNE) levels were measured with a biochemical index determination kit. Cellular Reactive Oxygen Species (ROS) were quantified using flow cytometry. CETSA, pull-down, and co-immunoprecipitation (Co-IP) assays confirmed the interactions between GA and SLC7A11, as well as the ubiquitination promoted by SLC7A11. A xenograft mouse model was employed to validate the Anticancer effect of GA in vivo.

Results: We observed that GA significantly suppressed proliferation in human CRC cells. Additionally, GA treatment inhibited the migration, invasion, and colony formation of CRC cells. Subsequently, through the use of specific inhibitors, we discovered that the suppression of CRC cells by GA was dependent on Ferroptosis rather than Autophagy or Apoptosis. Previous research has demonstrated that GA treatment significantly triggers Ferroptosis. Mechanistically, GA treatment promotes the degradation of the SLC7A11 protein, which plays a crucial role in Ferroptosis. Notably, the knockdown of SLC7A11 abolished the detrimental effects of GA on the proliferation of CRC cells and reversed GA-induced Ferroptosis in CRC cells both in vivo and in vitro. Further research has shown that GA can directly bind to the SLC7A11 protein and promote its ubiquitination.

Conclusion: Our research provides compelling evidence that GA may serve as a potential agent for suppressing the progression of CRC by inducing Ferroptosis and promoting the ubiquitination and degradation of SLC7A11.

Colorectal cancer; Ferroptosis; Gingerenone A; SLC7A11; Ubiquitination.