Fig. 8
The combination of ERO1α inhibition and IKE exerts an effective inhibitory effect on the growth of PDO and PDX models. A Schematic workflow of the generation of LSCC organoids and PDX models. B Representative IHC images of CK13, p63, Ki-67, p-S6, and ERO1α staining from LSCC tissues and organoids. Scale bar, 40 μm. C LSCC organoids were infected with lentiviruses expressing shRNAs targeting ERO1α (shERO1α1) or a non-targeting shRNA (shSc). The level of ERO1α was detected by western blotting. D shERO1α.1 or shSc lentiviruses-infected organoids were treated with IKE (50 μM) or DMSO for 24 h. The cell viability of organoids was determined by Cell-Titer Glo-3D cell viability assay. Left panels: representative phase contrast images. Right panels: quantitation of the data. Scale bar, 50 μm. E and F The expression of p-S6 and ERO1α in the PDX tumor tissues, primary tumor tissues and ANM tissues were analyzed by IHC (E) and western blotting (F). G–I Tumor images (G), tumor volume (H), and tumor weight (I) of PDX model tumors treated with ERO1α siRNAs or siNC, together with IKE (40 mg/kg) or vehicle. n = 5 mice per group. J IHC staining of PDX tumor tissues using the indicated antibodies. Scale bar, 40 μm. K MDA assay was used to detected lipid peroxidation levels in randomly selected PDX tumor section. L Body weight of mice. M Schematic illustration of the activated ERO1α/IL-6/STAT3/SLC7A11 pathway is critical for mTORC1-mediated ferroptosis resistance and tumor progression. Error bars indicate mean ± SD of triplicate (if mentioned otherwise) samples. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001