Marine-derived peptides from Rapana venosa inhibit breast cancer cell growth through synergistic mechanisms with doxorubicin.

Marine natural products are a promising source of novel anticancer agents. This study evaluated the antitumor activity of peptide fractions derived from the marine gastropod Rapana venosa against human breast cancer cell lines MCF-7 (estrogen receptor-positive) and MDA-MB-231 (triple-negative). Peptides were isolated by enzymatic hydrolysis and Fast Protein Liquid Chromatography. All seven isolated fractions were systematically screened against four human cancer cell lines (MCF-7, MDA-MB-231, CaCo-2, and HepG2) to assess cancer type selectivity. Cytotoxicity was assessed using MTT assay, while cell cycle progression and apoptosis were analyzed by flow cytometry. Gene expression changes were examined by RT-qPCR. Two peptide fractions, RV1 and RV2, demonstrated remarkable selectivity for breast cancer cells, exhibiting 25-95-fold higher potency compared to other cancer types. These fractions showed significant dose-dependent cytotoxicity with IC values of 6.887-7.288 μg/ml (RV1) and 4.886-6.268 μg/ml (RV2) against breast cancer cells. Both fractions induced G0/G1 cell cycle arrest and promoted apoptosis through multiple pathways, upregulating pro-apoptotic genes (TP53, AIFM1, CASP3, BAX) and downregulating anti-apoptotic markers (BCL2, miR-155). Most significantly, combination treatments with doxorubicin resulted in remarkable synergistic effects, with RV2 + doxorubicin achieving 78.9% total apoptosis compared to 33.5% with doxorubicin alone. These results indicate that R. venosa-derived peptides exert selective anticancer effects against breast cancer cells through multiple mechanisms. The observed selectivity and synergism with doxorubicin suggest their potential as targeted adjuvant agents in combination chemotherapy. Further structural characterization and in vivo studies are needed to advance their therapeutic development.