Since ferroptosis was first described in 2012, it has attracted considerable attention in the medical community as an emerging mode of cell death driven by iron-dependent lipid peroxidation, distinct from apoptosis, necrosis, and autophagy. This unique cell death mode is regulated by multiple pathways, including the classical System Xc–glutathione peroxidase 4 (GPX4) and ferroptosis suppressor protein 1 (FSP1)-CoQ10 systems, as well as lipid metabolism, mitochondrial metabolism, cholesterol metabolism, sex hormone metabolism, and other auxiliary pathways. Notably, the cholesterol biosynthesis pathway exhibits bidirectional regulation—7-dehydrocholesterol (7-DHC) acts as a “sacrificial antioxidant” to inhibit ferroptosis, whereas elevated cholesterol levels promote it. Sex hormones regulate MBOAT1/2 to remodel membrane phospholipids and reduce ferroptosis susceptibility. Key signaling pathways, such as P53/SLC7A11, Nrf2-Keap1, and Hippo-YAP/TAZ, further fine-tune ferroptosis through iron homeostasis and redox balance regulation. Crosstalk exists between ferroptosis and other cell death modalities: copper chelators sensitize cells to ferroptosis, while lipid peroxidation products activate pyroptosis via the NLRP3 inflammasome. Novel inhibitors, including JKE-1674 (FSP1-targeted) and SRS11-92 (lipid radical scavenger), show improved pharmacokinetics and tissue specificity, while nanoparticle-based delivery systems enhance targeting efficacy. Multiple cancer cells are highly susceptible to ferroptosis, and ferroptosis is implicated in neurodegenerative diseases, ischemia-reperfusion injury, and metabolic disorders. This review systematically summarizes the molecular mechanisms, regulatory networks, novel modulators, and clinical translation progress of ferroptosis, highlighting its potential as a therapeutic target for various diseases and providing insights for future research and clinical application.