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Updated: May 16, 2026
Revealing the Ferroptotic Phenotype of Medulloblastoma
Published on: March 15, 2024
Discovery of ferroptosis drugs targeting SLC7A11/BECN1 based on living cell quantitative FRET imaging
Zhiyu Xiao1, Ziru Wu1, Miao Tan1
1MOE Key Laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Laser Life Science, College of Biophotonics, School of Optoelectronic Science and Engineering, South China Normal University, Guangzhou, 510631, China.
Abstract:
This study aims to establish a method to discover ferroptosis drugs targeting the SLC7A11/BECN1 interaction based on live-cell quantitative fluorescence resonance energy transfer (FRET) imaging. In MDA-MB-231 cells, Imidazole Ketone Erastin (IKE) treatment induced ferroptosis verified by an 80% increase in Fe2+, a 38% increase in MDA, and a 53% decrease in GSH. Western blotting, protein co-localization analysis, and quantitative FRET microscopy collectively demonstrated that IKE promoted cytoplasmic interaction between SLC7A11 and BECN1 in living cells, resulting in decreased SLC7A11 protein levels, indirect inhibition of GPX4 activity, and ultimately ferroptosis induction. Gene manipulation experiments confirmed that SLC7A11 inhibited IKE-induced ferroptosis while BECN1 promoted IKE-induced ferroptosis. These results indicate that the interaction between SLC7A11 and BECN1 represents a viable target for ferroptosis regulation. Five drugs (IKE, SOR, LAP, LIN, ML162) were used for FRET drug screening. In MDA-MB-231 cells, IKE, SOR and LAP significantly promoted the interaction between SLC7A11 and BECN1 (EDmax was 0.47 ± 0.02, 0.34 ± 0.02, 0.30 ± 0.03, respectively) compared with the control group (EDmax was 0.16 ± 0.01), while LIN and ML162 had no effect (EDmax was 0.19 ± 0.02, 0.14 ± 0.01, respectively). In MCF-7 cells, the EDmax values of the control, IKE-, SOR-, LAP-, LIN-, and ML162-treated cells were 0.17 ± 0.04, 0.40 ± 0.01, 0.31 ± 0.01, 0.21 ± 0.01, 0.17 ± 0.01, 0.17 ± 0.02, respectively, consistent with the previous results, indicating the accuracy and effectiveness of this method. In summary, the SLC7A11/BECN1 interaction-based FRET imaging provides a quantitative method for the discovery of ferroptosis drugs in living cells.
Insights
This study introduces a live-cell imaging method to find ferroptosis drugs targeting the SLC7A11/BECN1 interaction. The method successfully identified drugs like IKE, SOR, and LAP that modulate this interaction, aiding ferroptosis drug discovery.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Ferroptosis is a regulated form of cell death implicated in various diseases.
- Targeting specific protein-protein interactions is a promising strategy for drug discovery.
- The interaction between solute carrier family 7 member 11 (SLC7A11) and BCL2-associated autophagy regulator (BECN1) is a potential target for ferroptosis modulation.
Purpose of the Study:
- To establish a quantitative live-cell imaging method using fluorescence resonance energy transfer (FRET) to detect the SLC7A11/BECN1 interaction.
- To utilize this FRET-based method for screening and identifying potential ferroptosis-inducing drugs.
- To validate the identified drugs' effects on the SLC7A11/BECN1 interaction and ferroptosis induction.
Main Methods:
- Quantitative live-cell fluorescence resonance energy transfer (FRET) imaging was employed to monitor the SLC7A11/BECN1 interaction in real-time.
- MDA-MB-231 and MCF-7 cell lines were used to validate the FRET method and drug screening.
- Western blotting and co-localization analysis were performed to confirm protein interactions and ferroptosis markers (Fe2+, MDA, GSH, GPX4 activity).
Main Results:
- The FRET method successfully quantified the SLC7A11/BECN1 interaction in living cells.
- Imidazole Ketone Erastin (IKE) treatment induced ferroptosis and promoted the cytoplasmic interaction between SLC7A11 and BECN1.
- Drug screening identified IKE, SOR, and LAP as compounds that significantly promote the SLC7A11/BECN1 interaction, indicating their potential as ferroptosis inducers.
Conclusions:
- The SLC7A11/BECN1 interaction is a viable target for ferroptosis regulation.
- The developed FRET-based imaging assay provides a quantitative and effective platform for discovering ferroptosis drugs.
- This method facilitates the identification of novel therapeutic agents targeting ferroptosis by monitoring specific protein-protein interactions in live cells.
