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Arenobufagin causes ferroptosis in human gastric cancer cells by increasing rev-erbα expression
Ke Chen1,2, Angling Li3, Jian Wang3
1Department of General Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, PR China.
Background And Aims:
Gastric cancer is the fifth most diagnosed malignant tumor worldwide with limited effective chemotherapy. Ferroptosis is a new type of programmed cell death, which is becoming as a novel therapeutic target for tumors. Arenobufagin (ArBu) is a bufadienolide isolated from toad skin and venom, which exhibits broad-spectrum anti-tumor activity. It is unclear whether ArBu causes ferroptosis, thereby exhibiting anti-tumor activity in gastric cancer. We aimed to determine whether ArBu causes ferroptosis in cultured human gastric cancer cells.
Experimental Procedure:
Different human gastric cancer cells were treated with ArBu (5-20 μM, 48 h). Indicators of apoptosis and ferroptosis were measured. CRISPR/Cas-9 system was employed to delete Nr1d1 gene.
Results:
ArBu incubation reduced cell viability in a concentration-dependent manner. ArBu caused ferroptosis but not apoptosis at a lower concentration (10 μM), despite it caused both of them at a higher concentration (20 μM). Cotreatment with a selective ferroptosis inhibitor ferrostatin-1 protected against ArBu (10 μM)-induced reduction in cell viability. ArBu-mediated ferroptosis was associated with abnormal expression of genes involved in iron uptake, lipid peroxidation, and antioxidants. Particularly, Nr1d1 gene expression was most significantly increased after ArBu treatment. Furthermore, activating Rev-erbα encoded by Nr1d1 by a selective agonist GSK4112 (1 and 2 μM, 48 h) caused ferroptosis. In contrast, Rev-erbα knockout using the CRISPR/Cas-9 system diminished ArBu-induced ferroptosis in cultured human gastric cancer cells.
Conclusion:
ArBu causes ferroptosis by increasing Rev-erbα expression in human gastric cancer cells. This has implications of ArBu as a promising therapy for gastric cancer.
Section:
1. Natural Products.
Taxonomy Classification By Evise:
Traditional medicine, pharmacology, gastric cancer, signal pathway.
Insights
Arenobufagin (ArBu) induces ferroptosis, a novel cell death pathway, in gastric cancer cells by upregulating Rev-erbα. This finding suggests ArBu as a potential therapeutic agent for gastric cancer treatment.
Area of Science:
- Oncology
- Cell Biology
- Pharmacology
Background:
- Gastric cancer is a leading cause of cancer death with limited chemotherapy options.
- Ferroptosis, a distinct form of programmed cell death, presents a novel therapeutic target for cancers.
- Arenobufagin (ArBu), a bufadienolide from toad venom, shows broad-spectrum anti-tumor activity, but its mechanism in gastric cancer is unknown.
Purpose of the Study:
- To investigate whether Arenobufagin (ArBu) induces ferroptosis in human gastric cancer cells.
- To elucidate the role of Rev-erbα in ArBu-mediated ferroptosis.
Main Methods:
- Human gastric cancer cells were treated with varying concentrations of ArBu.
- Apoptosis and ferroptosis indicators were assessed.
- Gene expression, including Nr1d1, was analyzed.
- CRISPR/Cas-9 was used for gene knockout studies.
- Ferroptosis was evaluated using a specific inhibitor (ferrostatin-1) and an agonist (GSK4112).
Main Results:
- ArBu reduced gastric cancer cell viability in a dose-dependent manner.
- ArBu induced ferroptosis, not apoptosis, at lower concentrations (10 μM), and both at higher concentrations (20 μM).
- Ferrostatin-1 protected cells from ArBu-induced cell death, confirming ferroptosis.
- ArBu treatment significantly increased Nr1d1 gene expression, which encodes Rev-erbα.
- Activating Rev-erbα induced ferroptosis, while its knockout diminished ArBu-induced ferroptosis.
Conclusions:
- Arenobufagin (ArBu) triggers ferroptosis in human gastric cancer cells.
- The mechanism involves the upregulation of Rev-erbα expression.
- ArBu demonstrates potential as a novel therapeutic strategy for gastric cancer.
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