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

Ruihua Duan; Jiao Dong; Mingjie Chen; Caixia Huang; Shenglong Gan; Runcong Liu; Ronghuan Yang; Hao Li; Zhoulei Li; Yisheng Lin; Ruihua Duan; Jiao Dong; Mingjie Chen; Caixia Huang; Shenglong Gan; Runcong Liu; Ronghuan Yang; Hao Li; Zhoulei Li; Yisheng Lin

doi:10.3934/molsci.2026005

AIMS Molecular Science

2026, Volume 13, Issue 1: 80-96. doi: 10.3934/molsci.2026005

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Research article Topical Sections

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

1.
Department of Medical Imaging, Maternal and Child Health Hospital of Hubei Province, Wuhan, Hubei 430070, China
2.
South China Academy of Advanced Optoelectronics, South China Normal University, Guangzhou, Guangdong 510006, China
3.
Department of Radiology, The First Affiliated hospital, Sun Yat-Sen University, 58 Zhongshan II Road, Guangzhou, Guangdong 510080, China
4.
Department of Interventional Radiology, Huizhou Central People's Hospital, Huizhou Guangdong 516001, China
5.
Shenzhen Medical Academy of Research and Translation, Shenzhen Guangdong 518107, China
6.
Department of Radiology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China
7.
Guangzhou Xinhua University, Guangzhou 510520, China
^# These four authors contributed equally to this work.

Received: 13 October 2025 Revised: 28 January 2026 Accepted: 02 February 2026 Published: 03 March 2026

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.
- molecular targeted therapy,
- apoptosis,
- cell cycle,
- liposomes,
- hepatocellular carcinoma,
- nanomedicine,
- cantharidin
Citation: Ruihua Duan, Jiao Dong, Mingjie Chen, Caixia Huang, Shenglong Gan, Runcong Liu, Ronghuan Yang, Hao Li, Zhoulei Li, Yisheng Lin. Molecular mechanisms of enhanced apoptosis in hepatocellular carcinoma via cantharidin-loaded liposomes[J]. AIMS Molecular Science, 2026, 13(1): 80-96. doi: 10.3934/molsci.2026005

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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.

Acknowledgments

This research was funded by The Natural Science Foundation of Guangdong Province, China (2019A1515012118, 2023A1515010388, 2025A1515012376) and Scientific research capacity improvement project of key construction disciplines in Guangdong Province, China (2024ZDJS129) and Huizhou Science and Technology Project, China (2022CZ010414).

Conflict of interest

The authors declare no conflicts of interest in this paper.

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