The anticancer and antioxidant activities from sea cucumber (Stichopus variegatus) flour dried vacuum oven have not been widely reported. This study aimed to determine in vitro inhibition of enzyme and water extracts from sea cucumber flour against colon cancer cells WiDr, breast cancer cells T47D, and normal cells Vero, then the apoptotic induction. Sea cucumber flour was made by vacuum oven. It was water extracted and hydrolyzed by digestive enzyme gradually. The water extract (SV-WE) and enzyme hydrolysis (SV-EE) were tested for their antioxidant activities using DPPH method. Samples of SV-WE and SV-EE were tested for their anticancer activities using MTT assays. Antioxidant activity of SV-EE at 1.67 ± 0.05 mg/mL was significantly different from SV-WE at 2.30 ± 0.30 mg/mL. SV-EE and SV-WE have anticancer activity against WiDr cells at 13.01 ± 2.75 μg/mL and 69.37 ± 24.25 µg/mL, respectively. The IC50 value of apoptotic ability of cell inductions from SV-EE was 64.9 ± 1.63%. SV-WE and SV-EE showed higher anticancer activity against WiDr cells and T47D cells. Gamma sea cucumber flour from Indonesia could be utilized as a potential ingredient in functional foods.