The exogenous enzymatic preparation process of Brassica rapa L. isothiocyanate (BRIe) was optimized,and anti-cervical cancer activity of the enzymatic products were analyzed and evaluated.The exogenous myrosinase was used for the degradation of B. rapa glucosinolate and the optimal preparation process for the enzymatic method was determined by one-way and orthogonal optimization experiments:the amount of myrosinase addition was 20 μL,enzymatic temperature was 70 ℃,pH 6.5,enzyme time was 2 h,and the ascorbic acid concentration was 0.01 mg/g.Under these conditions,the extraction rate of isothiocyanate was 11.13%.There are seven chemical components of the enzymatic hydrolysis products obtained by GC-MS identification and analysis,namely 3-amino-2-oxazolidinone,2-oxazolidinethione,2-oxazolidinethione,3-butenyl isothiocyanate,2- Phenethyl isothiocyanate,isopropyl isothiocyanate,2-phenylethyl isothiocyanate.The proliferation inhibitory activity of BRIe on cervical cancer Hela,Siha and U14 cells was detected by MTT assay,in which BRIe had the strongest inhibitory effect on Hela cells,with a 72 h IC₅₀ of 97.48 μg/mL;Cell scratch experiments showed that BRIe inhibited the migration of Hela cells,PI flow cytometry staining exhibited that BRIe arrested the cell cycle in the S phase and G2/M phase,and Hoechst 33258,Annexin V-FITC and PI staining demonstrated that BRIe induced apoptosis in Hela cells.The findings provide a scientific basis for the development and utilization of B. rapa for functional foods and anti-cervical cancer drugs.