GDF10 Regulates Iron-Dependent Lipid Oxidation in Colorectal Cancer Cells Through Interaction With IGF2.

Abstract

Background: This study aims to identify genes with a high likelihood of genetic variation, significant expression differences, and prognostic implications in colorectal cancer (CRC) patients. The research also seeks to investigate the role and mechanisms of key genes in CRC through the analysis of health information data and a combination of internal and external experiments.

Methods: The sequence information of CRC patients was obtained from the Cancer Genome Atlas Program (TCGA) database. Key genes were identified through differential expression analysis, enrichment analysis, interaction analysis, and survival curve analysis. Subsequently, Growth and Differentiation Factor 10 (GDF10) overexpression and knockout cell models were developed to investigate the impact of GDF10 on CRC cells using assays such as CCK8, flow cytometry, Transwell, and subcutaneous tumor formation in nude mice. Mechanistically, the effects of GDF10 and IGF2 genes on autophagy, apoptosis, and ferroptosis were studied by introducing different death pathway inhibitors. Changes in reactive oxygen species (ROS), GSH, and MDA levels in cells following silencing were also assessed. Furthermore, the influence of GDF10 and its interacting protein IGF2 on ferroptosis was evaluated through ELISA and fluorescence staining.

Results: The analysis of patients in the TCGA database revealed that GDF10 expression was low in microsatellite stable (MSS) type and high in microsatellite instability (MSI) type. It was found to interact with IGF2, with low expression associated with a better prognosis. Overexpression of GDF10 was shown to significantly enhance the proliferation and invasion abilities of CRC cells, whereas low expression promoted cell apoptosis. Within the pathways of autophagy, apoptosis, and ferroptosis, low expression of GDF10 primarily regulated ferroptosis in cells. This regulation involved promoting the iron-dependent lipid oxidation process by mediating binding with IGF2, leading to increased concentrations of iron ions and oxidative metabolites in cells. This process also reduced the formation of lipid droplets and inhibited tumor development.

Conclusion: GDF10 plays a crucial role in regulating ferroptosis in CRC cells through mediating IGF2 interaction, suggesting it as a promising therapeutic target for CRC.