Abstract 2828: Anthocyanin-induced autophagosome accumulation and mitochondrial dysfunction drive apoptosis in liver cancer cells

Abstract Introduction: Liver cancer remains one of the most lethal malignancies worldwide. Hepatocellular carcinoma (HCC) accounts for most cases. Existing therapies provide limited efficacy, especially in advanced stages, highlighting the urgent need for alternative treatments. Recent studies have demonstrated that anthocyanins, a natural plant-derived compound, show anticancer potential. Dracorhodin Perchlorate (DP), a synthetic analog of anthocyanin found in Daemonorops draco fruit, exhibits cytotoxic effects in cancer cells. This study investigates the mechanisms underlying DP’s cytotoxicity in HCC. Methods: We examined the effect of DP on human liver cancer cells (SK-HEP1). We employed an integrative experimental approach combining cell viability assay, death pathway analysis, autophagy monitoring, lysosomal function evaluation, oxidative stress assessment, and mitochondrial integrity. Western blot, immunocytochemistry, and live cell imaging were employed. Results: Our result shows that DP significantly reduced SK-HEP1cell viability in a time- and dose-dependent manner. The IC50 values decreased from 60 µM at 24 hours, 26 µM at 48 hours, and 18 µM at 72 hours, reaching 12 µM at 96 hours. We observed decreased pro-caspase-3 levels in DP-treated cells, confirming apoptosis. We found no change in phosphorylated MLKL levels, ruling out necroptosis. Interestingly, we observed a significant increase in LC3II/I ratios and LC3 puncta, indicating autophagosome accumulation. However, we noted no increase in lysosomal activity-based DQ-BSA and Lysotracker assays, suggesting autophagic flux inhibition. Furthermore, we detected a significant rise in reactive oxygen species (ROS) levels following DP treatment, accompanied by elevated HO-1 expression. We found evidence of mitochondrial dysfunction, as Mitotracker assays revealed fragmented mitochondria and reduced cytochrome c release. We observed that Bafilomycin A1 amplified DP-induced apoptosis, supporting the role of autophagic flux inhibition in driving cell death. Conclusions: Taken together, these findings reveal that DP induces apoptosis in liver cancer cells by driving autophagosome accumulation, oxidative stress, and mitochondrial damage. Our work suggests that DP holds potential as an anthocyanin-based therapy for liver cancer. Citation Format: Anyssa Rodriguez, Samantha Tamayo, Aarya Tripathi, Sumeet Chauhan, Subash Chauhan, Nirakar Sahoo. Anthocyanin-induced autophagosome accumulation and mitochondrial dysfunction drive apoptosis in liver cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2828.