Abnormal upregulation of the intracellular antioxidant Glutathione (GSH) plays a role in promoting tumor proliferation, inducing drug resistance and inhibiting ferroptosis in various malignant tumors, including hepatocellular carcinoma [HCC].
Targeting the upregulation mechanism of glutathione in HCC may become a therapeutic strategy to improve the prognosis of patients.
On August 21, 2025, Yuan Yufeng and Chen Xi from Wuhan University Zhang Zhonglin and Wenzhi He co-corresponds. published online in Cancer Research (IF=12.5) titled "The Acetyltransferase ARD1 Induces Glutathione Synthesis to Facilitate Research paper on "Ferroptosis Evasion in Hepatocellular Carcinoma".
This study, through genome-wide CRISPR-Cas9 screening and targeted metabolomics analysis, identified acetyltransferase ARD1 as a key promoter of de voo glutathione synthesis in HCC.
It is worth noting that the upregulation of ARD1 is positively correlated with elevated glutathione levels and poor prognosis in HCC patients.
In vitro and in vivo functional experiments have shown that ARD1 promotes the proliferation of HCC cells and inhibits ferroptosis in a glutathione-dependent manner.
The stable isotope labeling technique based on LC-MS /MS shows that ARD1 increases glutathione content by stabilizing the mRNA level of the γ -glutamyl cysteine ligase catalytic subunit (GCLC), a process mediated by the RNA-binding protein PABPC1.
Mechanistically, ARD1 mediates the acetylation of lysine at position 167 of PABPC1, enhancing its retention in the cytoplasm by disrupting the formation of the complex between PABPC1 and importin α7.
PABPC1 in the cytoplasm then interacts with eIF4G, synergistically stabilizing GCLC mRNA, preventing its degradation, increasing glutathione synthesis, and ultimately endowing HCC cells with ferroptosis resistance.
Hydrogen peroxy-induced oxidative stress can inhibit the ubiquitination and degradation of ARD1, thereby promoting the nuclear translocation of PABPC1 and inducing the expression of GCLC.
In the transplantation tumor model derived from HCC patients with high expression of ARD1 and GCLC, inhibition of ARD1 can enhance sorafenib's ferroptosis.
This study reveals an oxidative stress-ARD1-PABPC1-GCLC axis, which plays a key role in glutathione metabolic reprogramming and ferroptosis regulation in HCC, and provides a strategic basis for ferroptosion-based targeted therapy of HCC.
Hepatocellular carcinoma (HCC) accounts for 80% of primary liver cancers and is the sixth most commonly diagnosed cancer and the third leading cause of cancer-related deaths worldwide.
Research predicts that by 2040, the global annual incidence of liver cancer will increase by 55.0%, with an estimated 1.4 million new HCC cases and 1.3 million deaths.
As most HCC patients are diagnosed at an advanced stage and have a high postoperative recurrence rate (with a 5-year recurrence rate of 50% to 70%), the therapeutic effect of radical surgery is often not satisfactory.
Despite significant progress made in various treatment strategies, including immune checkpoint blockade (ICB) therapy, the 5-year overall survival rate of HCC patients remains below 20%.
A thorough understanding of the pathogenesis of HCC is crucial for identifying molecular targets for diagnosis and treatment.
Glutathione (GSH) is an endogenous antioxidant in cells that plays a core role in regulating key signaling pathways such as cell differentiation, proliferation, apoptosis, ferroptosis and immune function.
Glutathione is catalyzed and synthesized by cysteine, glutamic acid and glycine through two rate-limiting enzymes - glutamate-cysteine ligase (GCL) and glutathione synthase (GSS).
The cysteine residues in the newly synthesized Glutathione can be oxidized to oxidized glutathione (GSSG) by non-enzymatic free radicals, reactive oxygen species (ROS), or reactive nitrogen species (RNS), thereby protecting cells from oxidative damage.
The activation of the transcription factor Nrf2 can promote the synthesis of Glutathione, assist glioma cells in resisting intracellular REDOX crises and inhibit apoptosis.
It is worth noting that the level of Glutathione in HCC tissues is abnormally elevated, and reducing Glutathione can enhance the sensitivity of HCC cells to ferroptosis and sorafenib treatment.
These findings highlight the crucial role of Glutathione metabolism in the occurrence, progression and drug resistance of various malignant tumors. However, the specific mechanism of Glutathione metabolic reprogramming in HCC remains unclear and awaits further study.
Elevated glutathione levels in tumor cells can alleviate DNA breaks, protein carbonization and lipid peroxidation caused by excessive oxidative damage.
Ferroptosis is a regulated form of cell death characterized by iron-dependent lipid peroxidation, involving a variety of neurodegenerative diseases, autoimmune diseases, ischemia-reperfusion injury and tumorigenesis, and shows significant potential for anti-tumor treatment.
Given the crucial role of glutathione in maintaining cellular REDOX homeostasis, its metabolic abnormalities are closely related to susceptibility to ferroptosis and drug resistance.
For instance, ACTL6A promotes glutathione synthesis by inducing GCLC expression, thereby reducing the sensitivity of gastric cancer cells to ferroptosis.
The TRIM34-UPF1 complex can stabilize GPX4 and enhance the sensitivity of liver cancer cells to PD-1 treatment. The mechanism is related to the inhibition of ferroptosis by elevated glutathione levels.
These studies indicate that targeting glutathione metabolism to induce ferroptosis in tumor cells holds significant therapeutic potential.
In this study, through genome-wide CRISPR-Cas9 screening, acetyltransferase ARD1 was identified as an important regulatory factor for glutathione synthesis in HCC cells.
The author found that ARD1 promotes the proliferation of HCC cells and inhibits ferroptosis in a glutathione-dependent manner.
By using stable isotope tracer metabolic analysis (SIRM) based on LC-MS/MS and co-immunoprecipitation - tandem mass spectrometry technology, the authors revealed that ARD1 promotes de novo synthesis of glutathione by stabilizing GCLC mRNA.
Mechanistically, ARD1 acetylates the lysine at position 167 (K167) of PABPC1, preventing its nuclear translocation, and works in coordination with eIF4G to stabilize GCLC mRNA.
importantly, ARD1-mediated glutathione synthesis can be induced by exogenous oxidative stress.
The author's research results suggest that ARD1 is a potential therapeutic target for HCC and provide a new perspective for understanding glutathione metabolic reprogramming and ferroptosis regulatory networks in hepatocellular carcinoma.
