Objective: To study the best preparation technology of maize yellow pigment by ultrasonic assisted with enzymatic method, and the inhibitory effect of them on the growth of human gastric cancer cells (BGC-823). Methods: The optimum conditions of maize yellow pigment were extracted by ultrasonic assisted with enzymatic method by single factor and response surface experiments. The inhibition of maize yellow pigment on BGC-823 cells was detected by morphological observation and MTT assay to evaluate the antitumor activity. Results: the maize yellow pigment best extraction condition of ultrasonic with enzymatic method is: enzyme activity 10 000 U/g, ultrasonic power 400 W, enzymolysis temperature 35 ℃, the maximum extraction yield was (72.64 ± 0.42) μg/g under the condition, the maize yellow pigment has a certain inhibitory effect on BGC-823 cell were studied by cell morphological observation and MTT test. Conclusion: The extraction of maize yellow pigment by ultrasonic assisted with enzymatic method is a kind of effective extraction method, the maize yellow pigment prepared has strong inhibitory effect on BGC-823 cells.