Abstract A002: Punicalagin impedes malignant traits in ovarian adenocarcinoma cells through ROS-linked apoptosis and autophagy pathways

Abstract Ovarian epithelial adenocarcinoma remains a formidable adversary in women's health, often diagnosed late and managed with cytotoxic chemotherapies that jeopardize healthy tissues and compromise quality of life. As interest grows in phytochemicals for safer and targeted anticancer therapy, punicalagin, a potent ellagitannin extracted from pomegranate, emerges as a compelling candidate. In this study, we interrogated the therapeutic potential of punicalagin on two human ovarian cancer cell lines, OVCAR-3 and SKOV-3, focusing on its impact on proliferation, migration, and cell death mechanisms. A range of punicalagin concentrations (6.25–200 µM) was tested over 24, 48, and 72 hours to assess dose- and time-dependent responses. Punicalagin significantly curtailed cell proliferation and suppressed colony-forming efficiency. Migration assays, including wound healing and transwell, revealed a marked inhibition of cellular motility post-treatment. MitoSOX™ staining identified increased mitochondrial ROS generation, indicating oxidative stress as a contributor to punicalagin’s mode of action. Flow cytometry with Annexin V/PI staining confirmed the induction of apoptosis in both cell lines, while acridine orange staining detected autophagic vacuoles only in OVCAR-3 cells. Western blot analysis showed enhanced expression of BAX and E-cadherin, suppression of N-cadherin, and conversion of LC3-I to LC3-II in OVCAR-3, suggesting activation of both apoptotic and autophagic pathways. In conclusion, punicalagin disrupts ovarian cancer cell proliferation and migration while activating cell death pathways through oxidative stress. These findings position punicalagin as a promising, multi-targeted natural compound for potential integration into ovarian cancer therapy protocols. Citation Format: Zeeshan Ahmad Bhutta, Md. F. Kulyar, Kyung-Chul Choi. Punicalagin impedes malignant traits in ovarian adenocarcinoma cells through ROS-linked apoptosis and autophagy pathways [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Ovarian Cancer Research; 2025 Sep 19-21; Denver, CO. Philadelphia (PA): AACR; Cancer Res 2025;85(18_Suppl):Abstract nr A002.