The sea cucumber body wall was subjected to enzymatic hydrolysis using papain. The relationship between the enzyme concentration (1–5%w/wprotein weight) and hydrolysis time (60–360 min) and the degree of hydrolysis (DH), yield, antioxidant activities, and antiproliferative activity in a HepG2 liver cancer cell line was determined. The surface response methodology showed that the optimum conditions for the enzymatic hydrolysis of sea cucumber were a hydrolysis time of 360 min and 4.3% papain. Under these conditions, a 12.1% yield, 74.52% DH, 89.74% DPPH scavenging activity, 74.92% ABTS scavenging activity, 39.42% H2O2scavenging activity, 88.71% hydroxyl radical scavenging activity, and 9.89% HepG2 liver cancer cell viability were obtained. The hydrolysate was produced under optimum conditions and characterized in terms of its antiproliferative effect on the HepG2 liver cancer cell line.