Integrative bioinformatics and experimental evidence revealed that 5-FU-resistant CRC cells demonstrate significant upregulation of cholesterol metabolism regulators, including lectin-type oxidized LDL Receptor 1 (LOX1), cholesterol 25-hydroxylase (CH25H), and Cytochrome P450 Family 7 Subfamily B Member 1 (CYP7B1). The gene discussed is CH25H; the disease is colorectal carcinoma.