Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer.

Ferroptosis is an iron-dependent nonapoptotic form of cell death that links iron, lipid, and glutathione levels to a variety of disease-related activities. However, the characteristics of ferroptosis in cervical carcinoma (CC) are poorly understood. We acquired raw data on CC cohorts from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) database. Key genes were identified using differential gene expression analysis and intersected for further immune infiltration, transcription regulation, gene set variation analysis (GSVA), gene set enrichment analysis (GSEA), and drug sensitivity analysis. We also used immunohistochemical (IHC) staining to confirm the expression of important genes in cervical cancer tissue and their prognostic relevance. Finally, gene silencing and cell coculture experiments were used to verify the biological functional mechanism and its role in the tumor microenvironment (TME). Through bioinformatics analysis, we discovered that GCH1 and H1.2 are key ferroptosis-related molecules in cervical cancer. GCH1 and H1.2 could act as useful prognostic markers in cervical cancer, and in addition to their connection with the tumor microenvironment, the possible transcriptional regulatory network, hallmark pathways and chemotherapy sensitivity were also clarified. IHC of the tissue microarray (TMA) and immunofluorescence spatial distance evaluation revealed that GCH1 was more highly expressed in cervical cancer tissue than in paracarcinoma tissue. For patients with cervical cancer, higher GCH1 expression corresponded to a lower M2 cell proportion and a higher M1/M2 ratio as well as a greater GCH1-M2 distance. Silencing GCH1 in SiHa cells blocked the cell cycle, promoted apoptosis, and inhibited the migration and invasion abilities of the cells, possibly through the inhibition of the phosphorylated PI3K/AKT/mTOR pathway. Coculture of the cells with macrophages revealed that the silencing of GCH1 led to decreased expression of tumor necrosis factor (TNF), a biomarker of M1 macrophages. In this study, we performed a thorough investigation of ferroptosis-related genes and identified the functional complexity of GCH1 during tumorigenesis in cervical cancer.