Molecular mechanisms of enhanced apoptosis in hepatocellular carcinoma via cantharidin-loaded liposomes

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Abstract

Objective: Hepatocellular carcinoma (HCC) remains a molecularly complex malignancy with limited therapeutic options. Cantharidin (CTD), a natural terpenoid, exhibits potent antitumor activity but suffers from poor bioavailability and dose-limiting toxicity. This study aimed to develop CTD-loaded liposomes (CTD-Lips) and investigate their molecular mechanisms in enhancing apoptosis and cell cycle regulation in HCC. Methods: CTD-Lips were prepared using an optimized thin-film method and thoroughly characterized for size, encapsulation efficiency, and drug release profile. Cellular uptake was evaluated using coumarin-6-loaded nanoparticles in HepG2 cells. Cytotoxicity was assessed by CCK-8 assay, while apoptosis and cell cycle distribution were analyzed via flow cytometry. In vivo antitumor efficacy and systemic safety were evaluated in HepG2-xenografted nude mice compared to cisplatin, with comprehensive histopathological and biochemical analyses. Results: Optimized CTD-Lips showed nanoscale characteristics (120.80 nm) with high encapsulation efficiency (96.20%) and sustained release. Cellular uptake was significantly enhanced, leading to superior cytotoxicity (IC₅₀: 1.415 μg/mL vs. 4.971 μg/mL for free CTD) and induced S-phase cell cycle arrest. Molecular analysis revealed that CTD-Lips promoted apoptosis through enhanced DNA damage response. In vivo, CTD-Lips achieved 62.4% tumor growth inhibition, outperforming cisplatin, while maintaining systemic safety with no significant hematological or biochemical alterations. Conclusions: CTD-Lips represent a molecularly targeted nanoplatform that enhances CTD's antitumor efficacy through regulation of cell cycle progression and apoptosis pathways. This study provides molecular-level insights into CTD-Lip-mediated anticancer mechanisms, supporting its potential as a novel molecular-targeted therapy for HCC.

Keywords

molecular targeted therapy / apoptosis / cell cycle / liposomes / hepatocellular carcinoma / nanomedicine / cantharidin

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