BACKGROUND: The requirement for relevant and safe drugs for cancer treatment is considered a challenge. Recently, marine isolated compounds with various therapeutic targets have attracted many researchers. AIM: Isolation and identification of potential anticancer peptides from edible Callista chione soft tissues. METHODOLOGY: C. chione specimens were collected and peptides were extracted, purified with FPLC, and tested on normal (hepatocytes and VERO) and cancer (HepG2, and HT-29) cells. Bioactive fractions were tested by tandem mass spectrometry. RESULTS: Five different fractions were purified according to ionic charges and two fractions (4 and 5) showed a potent anticancer activity with a total anticancer score threshold of ≥ 0.5, and hydrophilicity mean of 1.75 that related to stability and solubility. The apoptotic and autophagy-related markers were significantly up-regulated in both HepG2 and HT-29 cells treated with IC(50) of bioactive peptides' fractions 4 and 5, explaining their underlying mechanism of action. CONCLUSION: Natural source peptides derived from the soft tissue of C. chione could be exploited for the treatment of cancers and a deep in silico study will be performed for further investigation and deep function identification.