GO:0110076 negative regulation of ferroptosis: Mechanisms, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0110076 (negative regulation of ferroptosis) describes any biological process that stops, prevents, or reduces the frequency, rate, or extent of ferroptosis, an iron-dependent form of regulated cell death.
• The GPX4-GSH axis is the central brake on ferroptosis; loss of GPX4 activity permits lipid peroxidation and ferroptotic death.
• Negative regulators such as STAT3, HLF, and P23 suppress ferroptosis in cancer cells, contributing to tumor growth and chemoresistance.
• Ferroptosis heterogeneity in triple-negative breast cancer reveals that negative regulation of ferroptosis can be targeted to improve immunotherapy combinations.
• Pharmacological or genetic inhibition of negative regulators (e.g., STAT3, SCD1) sensitizes tumors to ferroptosis and reverses chemoresistance.
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect causal roles of negative regulators of ferroptosis in disease.
Description
Ferroptosis is an iron-dependent form of regulated cell death driven by excessive lipid peroxidation, and its negative regulation is critical for maintaining cellular homeostasis and survival under stress. The Gene Ontology term GO:0110076, negative regulation of ferroptosis, encompasses any process that stops, prevents, or reduces the frequency, rate, or extent of ferroptosis. This term is increasingly relevant because dysregulated ferroptosis suppression contributes to cancer progression, chemoresistance, and immune evasion, while excessive ferroptosis is implicated in neurodegeneration and ischemia-reperfusion injury. Understanding the molecular players that negatively regulate ferroptosis is therefore essential for developing targeted therapies. Key negative regulators include the glutathione peroxidase GPX4, which detoxifies lipid peroxides and is considered the master brake on ferroptosis. Beyond GPX4, signaling axes such as STAT3-ferroptosis negative regulatory axis in gastric cancer and HLF-mediated tumor-macrophage crosstalk in triple-negative breast cancer illustrate the diversity of negative regulatory mechanisms. Recent studies have also identified P23 as a negative regulator that blocks GPX4 degradation via chaperone-mediated autophagy in non-small cell lung cancer. These findings underscore the importance of GO:0110076 in cancer biology and beyond. This article synthesizes authoritative QuickGO data and verified PubMed literature to provide a research-grade overview of negative regulation of ferroptosis, covering its definition, mechanisms, key genes, disease relevance, and experimental models including CRISPR-based approaches.
negative regulation of ferroptosis At A Glance
| GO ID | GO:0110076 |
|---|---|
| GO term | negative regulation of ferroptosis |
| Ontology | biological_process |
| Synonym | none |
| Major function | Suppression of iron-dependent lipid peroxidation and ferroptotic cell death |
| Key negative regulators | GPX4, STAT3, HLF, P23, SCD1, NCOA4 (context-dependent) |
| Disease relevance | Cancer chemoresistance, immunotherapy response, neurodegeneration, ischemia-reperfusion injury |
| Research methods | CRISPR KO/point mutation/knock-in/overexpression, lipid peroxidation assays, ferroptosis inducers (e.g., erastin, RSL3) |
What Is GO:0110076?
GO:0110076 (negative regulation of ferroptosis) is a biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of ferroptosis. In practical terms, it includes molecular mechanisms that inhibit iron-dependent lipid peroxidation, enhance antioxidant defense, or promote iron sequestration and storage, thereby protecting cells from ferroptotic death.
Why Is negative regulation of ferroptosis Important in Cell Biology?
Negative regulation of ferroptosis is important because it determines whether cells survive or die under oxidative stress and iron overload, directly influencing cancer progression, chemoresistance, and immune evasion. In triple-negative breast cancer, ferroptosis heterogeneity and negative regulatory mechanisms shape immunotherapy outcomes, making this process a therapeutic target. In gastric cancer, inhibition of the STAT3-ferroptosis negative regulatory axis suppresses tumor growth and alleviates chemoresistance. In non-small cell lung cancer, P23 acts as a negative regulator by blocking GPX4 degradation, promoting tumor cell survival. Conversely, excessive negative regulation may contribute to neurodegeneration by preventing clearance of damaged cells, as suggested by studies on mitochondrial calcium uniporter complex in neurons. Thus, understanding GO:0110076 is essential for developing strategies to modulate ferroptosis in disease.
• Determines cell fate under oxidative stress and iron overload.
• Drives chemoresistance in gastric cancer via STAT3-ferroptosis axis.
• Shapes immunotherapy response in triple-negative breast cancer.
• Promotes tumor growth through HLF-mediated macrophage crosstalk.
• Blocks GPX4 degradation via P23 in NSCLC, enhancing survival.
• Protects neurons from ferroptosis via mitochondrial calcium uniporter complex modulation.
• Involved in metabolic and redox homeostasis.
• Target for sensitizing tumors to ferroptosis inducers.
• Potential therapeutic avenue in neurodegeneration and ischemia.
• Requires CRISPR models to dissect causal roles of negative regulators.
What Happens During negative regulation of ferroptosis?
GPX4-mediated detoxification of lipid peroxides
In simple terms: GPX4 acts like a cellular antioxidant that neutralizes harmful lipid peroxides before they can destroy the cell membrane.
GPX4 is a glutathione-dependent enzyme that reduces lipid hydroperoxides to harmless alcohols, thereby preventing the propagation of lipid peroxidation and ferroptosis. Loss of GPX4 activity is a hallmark of ferroptosis sensitivity, and its negative regulation is central to GO:0110076. Pharmacological inhibition of GPX4 (e.g., RSL3) induces ferroptosis, confirming its role as a negative regulator.
STAT3 signaling axis
In simple terms: STAT3 acts as a brake on ferroptosis by promoting survival signals in cancer cells.
In gastric cancer, inhibition of the STAT3-ferroptosis negative regulatory axis suppresses tumor growth and alleviates chemoresistance, indicating that STAT3 negatively regulates ferroptosis. Targeting this axis may sensitize tumors to ferroptosis inducers.
HLF-mediated tumor-macrophage crosstalk
In simple terms: HLF helps tumor cells talk to macrophages to avoid ferroptosis and resist chemotherapy.
HLF regulates ferroptosis, development, and chemoresistance of triple-negative breast cancer by activating tumor cell-macrophage crosstalk, thereby negatively regulating ferroptosis. This highlights the role of tumor microenvironment interactions in GO:0110076.
P23 blocks GPX4 degradation via chaperone-mediated autophagy
In simple terms: P23 protects GPX4 from being broken down, keeping the ferroptosis brake engaged.
In non-small cell lung cancer, P23 acts as a negative regulator of ferroptosis by blocking GPX4 degradation via chaperone-mediated autophagy. This mechanism stabilizes GPX4 and suppresses ferroptosis.
SCD1-mediated lipogenesis and NCOA4-mediated ferritinophagy
In simple terms: SCD1 changes lipid composition to make cells less vulnerable, while NCOA4 controls iron release from ferritin.
Salidroside sensitizes triple-negative breast cancer to ferroptosis by modulating SCD1-mediated lipogenesis and NCOA4-mediated ferritinophagy, indicating that these pathways contribute to negative regulation of ferroptosis. SCD1 alters lipid saturation, while NCOA4 regulates iron availability, both impacting ferroptosis sensitivity.
Mitochondrial calcium uniporter complex modulation
In simple terms: Tuning mitochondrial calcium uptake can protect neurons from ferroptosis.
Negative modulation of the mitochondrial calcium uniporter complex protects neurons against ferroptosis, demonstrating a neuroprotective mechanism of negative regulation. This suggests that mitochondrial calcium handling is part of GO:0110076 in neuronal contexts.
Key Genes Involved in GO:0110076 negative regulation of ferroptosis
The following genes and proteins are established negative regulators of ferroptosis, supported by verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| GPX4 | Glutathione peroxidase that detoxifies lipid peroxides | Master negative regulator; target for ferroptosis induction |
| STAT3 | Transcription factor promoting survival and negative regulation of ferroptosis | Inhibition suppresses tumor growth and chemoresistance in gastric cancer |
| HLF | Regulates tumor-macrophage crosstalk and chemoresistance | Negative regulator in triple-negative breast cancer |
| P23 | Blocks GPX4 degradation via chaperone-mediated autophagy | Negative regulator in NSCLC |
| SCD1 | Stearoyl-CoA desaturase involved in lipogenesis | Modulates lipid composition to suppress ferroptosis |
| NCOA4 | Ferritinophagy receptor controlling iron release | Affects iron availability and ferroptosis sensitivity |
| SLC7A11 | Cystine/glutamate antiporter supporting glutathione synthesis | Indirect negative regulator by maintaining GSH levels |
| GCLC | Glutamate-cysteine ligase catalytic subunit for GSH synthesis | Supports GPX4 function and negative regulation |
| GCLM | Glutamate-cysteine ligase modifier subunit | Modulates GSH synthesis and ferroptosis resistance |
| FTH1 | Ferritin heavy chain for iron storage | Reduces labile iron, negatively regulating ferroptosis |
| FTL | Ferritin light chain for iron storage | Iron sequestration suppresses ferroptosis |
| NFS1 | Cysteine desulfurase involved in iron-sulfur cluster biogenesis | Supports redox homeostasis and negative regulation |
| CISD1 | Mitochondrial iron-sulfur protein | Protects against mitochondrial lipid peroxidation |
| ACSL4 | Acyl-CoA synthetase long-chain family member 4 | Promotes ferroptosis; its inhibition is negative regulation |
| LPCAT3 | Lysophosphatidylcholine acyltransferase 3 | Remodels phospholipids to influence ferroptosis sensitivity |
| NFE2L2 | Transcription factor regulating antioxidant response | Upregulates GPX4 and other antioxidants |
| MT1G | Metallothionein 1G | May modulate zinc/redox and ferroptosis |
How Is negative regulation of ferroptosis Regulated?
Negative regulation of ferroptosis is controlled by multiple signaling pathways. The STAT3-ferroptosis negative regulatory axis is a key mechanism in gastric cancer, where STAT3 activation suppresses ferroptosis and promotes chemoresistance. In triple-negative breast cancer, HLF activates tumor cell-macrophage crosstalk to negatively regulate ferroptosis. P23 blocks GPX4 degradation via chaperone-mediated autophagy, stabilizing GPX4 and suppressing ferroptosis in NSCLC. Additionally, SCD1-mediated lipogenesis and NCOA4-mediated ferritinophagy modulate ferroptosis sensitivity in breast cancer. Mitochondrial calcium uniporter complex activity also influences neuronal ferroptosis, with its negative modulation being protective. These pathways collectively fine-tune cellular susceptibility to ferroptosis.
negative regulation of ferroptosis and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| STAT3 | Gastric cancer chemoresistance | STAT3 knockout or point mutation in gastric cancer cell lines |
| HLF | Triple-negative breast cancer | HLF overexpression or knockout in TNBC cells |
| P23 | Non-small cell lung cancer | P23 knockout or overexpression in NSCLC cells |
| GPX4 | Ferroptosis sensitivity across cancers | GPX4 knockout or point mutation in cancer cell lines |
| SCD1 | Triple-negative breast cancer lipogenesis | SCD1 knockout or overexpression in TNBC cells |
Cancer chemoresistance and immunotherapy
Negative regulation of ferroptosis contributes to chemoresistance in gastric cancer through the STAT3 axis. In triple-negative breast cancer, ferroptosis heterogeneity and negative regulators like HLF shape immunotherapy responses, and targeting these mechanisms can improve combination strategies. P23-mediated GPX4 stabilization in NSCLC promotes tumor survival, making it a potential therapeutic target.
Neurodegeneration and neuroprotection
Negative modulation of the mitochondrial calcium uniporter complex protects neurons against ferroptosis, suggesting that enhancing negative regulation may be beneficial in neurodegenerative conditions. However, excessive suppression of ferroptosis could impair clearance of damaged cells, highlighting the need for context-specific modulation.
Metabolic and redox homeostasis
Negative regulation of ferroptosis is intertwined with metabolic and redox homeostasis, as GPX4 and glutathione synthesis pathways maintain cellular antioxidant capacity. Disruption of these pathways can tip the balance toward ferroptotic cell death, linking GO:0110076 to metabolic disorders.
From negative regulation of ferroptosis-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does GPX4 loss sensitize cells to ferroptosis? | GPX4 knockout cell line |
| Does STAT3 inhibition reverse chemoresistance? | STAT3 knockout or point mutation in gastric cancer cells |
| Does P23 stabilize GPX4? | P23 overexpression and knockout in NSCLC cells |
| Does HLF mediate macrophage crosstalk? | HLF knockout in TNBC co-culture models |
| Does SCD1 modulate lipid composition? | SCD1 overexpression or knockout in TNBC cells |
| Does NCOA4 affect iron availability? | NCOA4 knockout or tagged knock-in in breast cancer cells |
How to Study the negative regulation of ferroptosis Process
| Method | What It Measures | Typical Application |
|---|---|---|
| C11-BODIPY staining | Lipid peroxidation | Ferroptosis detection |
| MDA assay | Malondialdehyde levels | Oxidative stress quantification |
| Cell viability assay | Cell survival under ferroptosis inducers | Screening for negative regulators |
| CRISPR knockout screen | Gene essentiality for ferroptosis resistance | Discovery of novel negative regulators |
| RNA-seq | Transcriptional changes | Pathway analysis after gene modulation |
| Proteomics | Protein expression and modifications | Identifying GPX4 stability regulators |
| Western blot | Protein levels (e.g., GPX4, P23) | Validation of negative regulators |
| Co-immunoprecipitation | Protein-protein interactions | Chaperone-mediated autophagy studies |
Lipid peroxidation and ferroptosis assays
Measuring lipid peroxidation using C11-BODIPY or malondialdehyde (MDA) assays is standard to assess ferroptosis and its negative regulation. Cell viability assays with ferroptosis inducers (e.g., erastin, RSL3) help quantify negative regulatory capacity.
CRISPR-based genetic screens
Genome-wide CRISPR knockout screens can identify novel negative regulators of ferroptosis by selecting for cells that survive ferroptosis induction. These screens are powerful for discovering genes like GPX4 and others.
Transcriptomics and proteomics
RNA-seq and proteomics can reveal changes in antioxidant pathways, iron metabolism, and lipid remodeling upon modulation of negative regulators. For example, SCD1 and NCOA4 expression changes were identified in breast cancer ferroptosis studies.
Imaging and biochemical assays
Live-cell imaging of lipid ROS and iron levels, combined with Western blot for GPX4 and other regulators, provides mechanistic insights. Chaperone-mediated autophagy can be monitored by co-immunoprecipitation and lysosomal inhibitors.
How CRISPR Can Be Used to Study GO:0110076 negative regulation of ferroptosis
Knockout
CRISPR knockout of negative regulators such as GPX4, STAT3, or P23 can sensitize cells to ferroptosis, confirming their roles in GO:0110076. For example, GPX4 knockout induces ferroptosis in various cancer cell lines.
Point Mutation
Point mutations can dissect specific domains or residues required for negative regulation, such as GPX4 catalytic site mutations that abolish its antioxidant activity. Similarly, STAT3 point mutations can clarify its ferroptosis-suppressive function.
Knock-in
Knock-in of tagged versions (e.g., GFP-P23) allows tracking of protein localization and stability, as shown for P23 in NSCLC. This helps understand how negative regulators interact with GPX4 and other components.
Overexpression
Overexpression of negative regulators like HLF or P23 can confer ferroptosis resistance and chemoresistance, providing gain-of-function evidence. This approach is useful for validating therapeutic targets.
How EDITGENE Supports negative regulation of ferroptosis Research
Researchers studying negative regulation of ferroptosis-related genes often need to determine whether a candidate gene is causally involved in suppressing ferroptotic cell death or merely correlated with survival. CRISPR-based models provide the gold standard for establishing causality, enabling precise genetic perturbations that reveal how specific genes contribute to GO:0110076.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of ferroptosis research.
Frequently Asked Questions About negative regulation of ferroptosis
What is negative regulation of ferroptosis (GO:0110076)?
It is any biological process that stops, prevents, or reduces the frequency, rate, or extent of ferroptosis, an iron-dependent form of regulated cell death.
What genes are involved in negative regulation of ferroptosis?
Key genes include GPX4, STAT3, HLF, P23, SCD1, and NCOA4, among others.
How does GPX4 negatively regulate ferroptosis?
GPX4 detoxifies lipid peroxides using glutathione, preventing lipid peroxidation and ferroptotic cell death.
What is the role of STAT3 in ferroptosis?
STAT3 acts as a negative regulator of ferroptosis; its inhibition suppresses tumor growth and alleviates chemoresistance in gastric cancer.
How does P23 regulate ferroptosis?
P23 blocks GPX4 degradation via chaperone-mediated autophagy, stabilizing GPX4 and suppressing ferroptosis in NSCLC.
What is the connection between ferroptosis and cancer immunotherapy?
Ferroptosis heterogeneity in triple-negative breast cancer influences immunotherapy responses, and targeting negative regulators can improve combination strategies.
Can CRISPR be used to study negative regulation of ferroptosis?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect causal roles of negative regulators.
What experimental models are used to study GO:0110076?
Common models include GPX4 knockout, STAT3 knockout, P23 overexpression, and HLF knockout cell lines, often combined with lipid peroxidation assays.
Why is negative regulation of ferroptosis important in neurodegeneration?
Negative modulation of the mitochondrial calcium uniporter complex protects neurons against ferroptosis, suggesting therapeutic potential.
What methods measure negative regulation of ferroptosis?
Methods include C11-BODIPY staining, MDA assays, cell viability with ferroptosis inducers, CRISPR screens, RNA-seq, and proteomics.
Conclusion
GO:0110076 (negative regulation of ferroptosis) is a critical biological process that governs cell survival under oxidative stress and iron overload. Key negative regulators such as GPX4, STAT3, HLF, and P23 have been implicated in cancer chemoresistance, immunotherapy response, and neuroprotection. Understanding these mechanisms through CRISPR-based models and advanced omics will accelerate the development of targeted therapies. EDITGENE provides comprehensive services to support this research.
References
- 1. Huang G et al.. 2025. Salidroside sensitizes Triple-negative breast cancer to ferroptosis by SCD1-mediated lipogenesis and NCOA4-mediated ferritinophagy.. J Adv Res 74:589-607 PMID: 39353532
- 2. Yang F et al.. 2023. Ferroptosis heterogeneity in triple-negative breast cancer reveals an innovative immunotherapy combination strategy.. Cell Metab 35(1):84-100.e8 PMID: 36257316
- 3. Ouyang S et al.. 2022. Inhibition of STAT3-ferroptosis negative regulatory axis suppresses tumor growth and alleviates chemoresistance in gastric cancer.. Redox Biol 52:102317 PMID: 35483272
- 4. Sharma A et al.. 2021. Positive and Negative Regulation of Ferroptosis and Its Role in Maintaining Metabolic and Redox Homeostasis.. Oxid Med Cell Longev 2021:9074206 PMID: 34007410
- 5. Marmolejo-Garza A et al.. 2023. Negative modulation of mitochondrial calcium uniporter complex protects neurons against ferroptosis.. Cell Death Dis 14(11):772 PMID: 38007529
- 6. Seibt TM et al.. 2019. Role of GPX4 in ferroptosis and its pharmacological implication.. Free Radic Biol Med 133:144-152 PMID: 30219704
- 7. Li H et al.. 2022. HLF regulates ferroptosis, development and chemoresistance of triple-negative breast cancer by activating tumor cell-macrophage crosstalk.. J Hematol Oncol 15(1):2 PMID: 34991659
- 8. Chen J et al.. 2025. P23 acts as a negative regulator of ferroptosis in NSCLC by blocking GPX4 degradation via chaperone-mediated autophagy.. Mol Cancer 24(1):234 PMID: 41039570
