Disulfiram/Copper Combination as a Potential Therapeutic Approach for Hepatocellular Carcinoma - Targeting the ATF3-Mitochondrial Cell Death Pathway

Sep 28, 2026

Uploaded by Oliver

Hepatocellular carcinoma (HCC) represents a major public health issue globally, necessitating urgent development of new therapies. This study evaluated the cytotoxic efficacy of disulfiram (DSF) and copper (Cu) in HCC by assessing mitochondrial function and apoptosis mechanisms. DSF/Cu treatment produced a significant dose-dependent reduction in HCC cell viability, with increased reactive oxygen species, reduced ATP production, and diminished mitochondrial membrane potential. Caspase-9 and caspase-3 activation indicated apoptosis, while DSF/Cu synergistically upregulated ATF3. In vivo mouse tumor experiments further confirmed inhibited tumor growth in the DSF/Cu group, supporting ATF3-dependent mitochondrial apoptosis as a promising therapeutic strategy.

LanguageEnglish

CategoryHealthcare

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Pages14

Reading Time35 min

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Abstract

Introduction

Materials and Methods

Cell culture

Ivyspring International Publisher Journal of Cancer 2026; 17(1): 117-130. doi: 10.7150/jca.113442 Research Paper Disulfiram/Copper Combination as a Potential Therapeutic Approach for Hepatocellular Carcinoma: Targeting the ATF3-Mitochondrial Cell Death Pathway Jing Cao1\#, Jing Deng1\#, Xinhua Li1\#, Yaqiong Chen2, Jialei Wang1, Yutian Chong1􀀍, Jiao Gong2􀀍, Bingliang Lin1􀀍 1. Department of Infectious Diseases, Key Laboratory of Liver Disease of Guangdong Province, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, China. 2. Department of Laboratory Medicine, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, China. \#These authors have made equal contributions. 􀀍 Corresponding authors: Bingliang Lin, [linbingl@mail.sysu.edu.cn](linbingl@mail.sysu.edu.cn), Jiao Gong, [gongjiao@alumni.sysu.edu.cn](gongjiao@alumni.sysu.edu.cn), and Yutian Chong, [chongyt@mail.sysu.edu.cn](chongyt@mail.sysu.edu.cn). © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License ([https://creativecommons.org/licenses/by/4.0/](https://creativecommons.org/licenses/by/4.0/)). See [https://ivyspring.com/terms](https://ivyspring.com/terms) for full terms and conditions. Received: 2025.03.09; Accepted: 2025.11.06; Published: 2026.01.01 Abstract Hepatocellular carcinoma (HCC) represents a major public health issue globally, necessitating the urgent development of new therapies. The therapeutic efficacy of disulfiram (DSF) and copper (Cu) in HCC was investigated in the present study, focusing on cytotoxicity, mitochondrial function, and apoptosis to clarify the mechanistic basis of this drug combination. Our findings revealed a significant, dose-dependent reduction in HCC cell viability with DSF/Cu treatment. Further investigation showed increased reactive oxygen species (ROS) levels, decreased adenosine triphosphate (ATP) production, and a decline in mitochondrial membrane potential (MMP). These events culminated in the activation of caspase-9 and caspase-3, key enzymes in the apoptotic pathway, leading to cell death. Mechanistically, DSF/Cu synergistically increased the expression of activating transcription factor 3 (ATF3), a known tumor suppressor, in HCC cells. In vivo studies using a mouse tumor model supported these findings, demonstrating significantly inhibited tumor growth in the DSF/Cu group compared with the control group. Overall, our study findings suggest that the DSF/Cu combination exhibits significant therapeutic potential against HCC by modulating the ATF3-dependent mitochondrial apoptosis pathway, a strategy that warrants further preclinical exploration. Keywords: disulfiram; hepatocellular carcinoma; mitochondrial damage; apoptosis; ATF3 Introduction Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer, accounting for 75%–86% of cases. As a major global health concern, it imposes a substantial strain on healthcare systems worldwide [1] . Despite advancements in therapeutic approaches, the prognosis for advanced-stage HCC remains suboptimal, necessitating the exploration of innovative and effective treatment strategies [2] . Despite considerable progress in liver cancer diagnostics and therapeutics, surgical resection, liver transplantation, and radiofrequency ablation remain the mainstay of treatment for early-stage HCC [3] . Due to the inherent difficulties in detecting early-stage disease, the majority of cases are diagnosed at advanced phases, where molecular targeted therapy and immunotherapy play critical roles. Nevertheless, due to challenges such as high drug resistance and frequent recurrence, the overall prognosis of this patient population remains poor [4] . Therefore, identifying novel therapeutic strategies is crucial for improving clinical outcomes. Indeed, the high costs, failure rates, and long testing periods associated with developing new medicines emphasize

FAQs

What was the main therapeutic focus of the DSF/Cu combination in hepatocellular carcinoma?

The study focused on whether disulfiram/copper (DSF/Cu) could kill HCC cells by targeting mitochondrial function and triggering apoptosis through the ATF3-dependent pathway.

How did DSF/Cu affect HCC cell viability and mitochondrial-related markers?

DSF/Cu significantly reduced HCC cell viability in a dose-dependent manner and increased reactive oxygen species while decreasing ATP production and mitochondrial membrane potential.

What evidence supports that DSF/Cu induces apoptosis in HCC?

The treatment activated caspase-9 and caspase-3, key apoptotic enzymes, and promoted mitochondrial damage and apoptosis in HCC cells, with supportive in vivo tumor growth inhibition in mice.

Sep 28, 2026

Uploaded by Oliver