Fig. 7
The mTORC1/ERO1α/IL-6/STAT3/SLC7A11 signaling pathway exists in human cancer. A and B LIU-LSC-1 cells were infected with lentivirus expressing shRNAs targeting Raptor (shRaptor1 and shRapator2) or a control shRNA (shSc). Cell lysates were subjected to immunoblotting with the indicated antibodies (A), ERO1α and SLC7A11 mRNA levels were detected by qRT–PCR (B). C 12 paired LSCC tissues and the corresponding ANM tissues were subjected to immunoblotting with the indicated antibodies. D Representative IHC images of p-S6, ERO1α, IL-6, p-STAT3, and SLC7A11 staining from the LSCC tissues and ANM tissues. Scale bar, 20 μm. E–I LIU-LSC-1 were infected with lentiviruses expressing shRNAs targeting ERO1α (shERO1α1 and shERO1α2) or a non-targeting shRNA (shSc). E The protein and mRNA levels of ERO1α and SLC7A11 were determined by western blotting and qRT–PCR. F and G CCK-8 (F) and colony formation (G) assays were performed to evaluate cell growth. H and I The pro-angiogenic effect of ERO1α was detected by tube formation (H) and CAM assays (I). Representative images (left panels) and quantifications (right panels) are shown. J Cell viability of indicated cells following treatment with erastin for 24 h. K Representative phase-contrast images of indicated cells were treated with erastin (15 μM) in the absence or presence of Lip-1 (1 μM). The corresponding phase contrast images are shown. Scale bar, 100 μm. L–N The indicated cells were treated with or without erastin (15 μM) for 24 h, and then intracellular MDA (L), L-ROS (M), and intracellular GSH (N) were measured. O–R Tumor images (O), tumor volume (P), and tumor weight (Q) of shERO1α.1 LIU-LSC-1 xenograft tumors (n = 5 mice/group) treated with Lip-1 (10 mg/kg) or vehicle. R The relative MDA levels of the indicated tumors were measured. Error bars indicate mean ± SD of triplicate (if mentioned otherwise) samples. **P < 0.01; ***P < 0.001; ****P < 0.0001