Fig. 7: c-MET/NRF2/GPX4 axis is a target for synthetic lethality in ARID1A-KO colorectal cancer cells. | Cell Death & Differentiation

Fig. 7: c-MET/NRF2/GPX4 axis is a target for synthetic lethality in ARID1A-KO colorectal cancer cells.

From: ARID1A loss enhances sensitivity to c-MET inhibition by dual targeting of GPX4 and iron homeostasis, inducing ferroptosis

Fig. 7

A RT-qPCR analysis of GPX4 mRNA level with PHA treatment in HCT116 cells. ANOVA P < 0.01. B Immunoblot analysis showing PHA treatment downregulates GPX4 level in HCT116, Lovo, and SW480 cells. C c-MET silencing induces GPX4 downregulation in HCT116, Lovo, and SW480 cells. Inhibition of GPX4 level by PHA treatment in HCT116 (D) and RKO (E) cells. F, G Overexpression of GPX4 reversed synthetic lethality induced by PHA treatment in HCT116 ARID1A-KO cells. HCT116 ARID1A-KO cells were treated with or without PHA and GPX4 expression plasmid. The cell viability was determined by Alamar Blue assay. ANOVA P value of <0.01. H co-IP showing ARID1A binds with NRF2 in HCT116 cells. I ChIP of GPX4 promoter in HCT116 and ARID1A-KO cells using anti-NRF2 antibody. IgG in each cell line was used as a normalization control. ANOVA P < 0.01. J Immunofluorescences showing that ARID1A loss attenuates NRF2 nuclear localization in HCT116 cells. Scale bars, 20 μm. K c-MET overexpression promotes NRF2 nuclear localization in ARID1A-KO cells. Immunofluorescences show that c-MET overexpression promotes NRF2 nuclear localization in ARID1A-KO cells. Scale bars, 20 μm. L c-MET overexpression promotes the binding between NRF2 and GPX4 promoter. ChIP of GPX4 promoter in ARID1A-KO cells using anti-NRF2 antibody with or without c-MET expression plasmid. IgG in each cell line was used as a normalization control. ANOVA P < 0.01. M ChIP of GPX4 promoter in HCT116 cells using anti-ARID1A antibody with or without PHA treatment. IgG in each cell line was used as a normalization control. ANOVA P < 0.01. N Working model of the synthetic lethality. In ARID1A-WT cells, ARID1A promotes c-MET transcription and cooperatively regulates NRF2 transcription factor functions with c-MET, thereby promoting GPX4 transcription. ARID1A inactivation downregulates c-MET and GPX4 levels, creating a cellular dependency on the residual activity of c-MET and GPX4. ARID1A inactivation also downregulates iron-exporting protein SLC40A1, increasing intracellular iron accumulation and lipid peroxidation. Pharmacological inhibition of c-MET (such as PHA665452) in ARID1A-deficient CRC cells diminishes GPX4 transcription, resulting in increased lipid peroxidation and ultimately inducing ferroptotic cell death. PLOOH a reactive phospholipid hydroperoxide, PLOH non-reactive alcohol, PL• a carbon-centered radical on a phospholipid chain.

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