Abstract: Cytochrome P450(CYP)1B1, as a key enzyme in extrahepatic metabolism, is characterized by tissue-specific high expression in various malignancies, while showing low or no expression in normal tissues. This enzyme promotes tumor occurrence and development through dual mechanisms: on one hand, it catalyzes the production of genotoxic 4-hydroxyestradiol, and the resulting oxidized quinone metabolites can directly damage DNA and induce genomic instability; on the other hand, it metabolically activates procarcinogens such as polycyclic aromatic hydrocarbons to form DNA adducts, which can induce mutations in oncogenes and promote cancer development. Polymorphisms in the CYP1B1 gene significantly affect individual cancer susceptibility and demonstrate heterogeneity across different races and cancer types. Furthermore, CYP1B1 mediates chemotherapy resistance through drug metabolism inactivation(such as paclitaxel) and regulation of signaling pathways including AKT/ERK and FAK/SRC, making it an important target for the prevention and treatment of tumor drug resistance. Research into its inhibitors has revealed that compounds such as flavonoids, chalcones, and stilbenes have potential to reverse drug resistance through competitive binding or signaling pathway intervention. This article reviews the multidimensional roles of CYP1B1 in tumor metabolic reprogramming, genetic susceptibility, and treatment resistance, while discussing future research directions based on multi-omics integration and precise targeting strategies, thus providing new insights for early cancer diagnosis, chemoprevention, and combination therapy.