The dysregulated mechanistic target of rapamycin complex 1 (mTORC1) signaling plays a critical role in ferroptosis resistance and tumorigenesis. However, the precise underlying mechanisms still need to be fully understood. Endoplasmic reticulum oxidoreductase 1 alpha (ERO1α) expression in mTORC1-activated mouse embryonic fibroblasts, cancer cells, and laryngeal squamous cell carcinoma (LSCC) clinical samples was examined by quantitative real-time PCR (qRT–PCR), western blotting, immunofluorescence (IF), and immunohistochemistry. Extensive in vitro and in vivo experiments were carried out to determine the role of ERO1α and its downstream target, member 11 of the solute carrier family 7 (SLC7A11), in mTORC1-mediated cell proliferation, angiogenesis, ferroptosis resistance, and tumor growth. The regulatory mechanism of ERO1α on SLC7A11 was investigated via RNA-sequencing, a cytokine array, an enzyme-linked immunosorbent assay, qRT–PCR, western blotting, IF, a luciferase reporter assay, and a chromatin immunoprecipitation assay. The combined therapeutic effect of ERO1α inhibition and the ferroptosis inducer imidazole ketone erastin (IKE) on mTORC1-activated cells was evaluated using cell line-derived xenografts, LSCC organoids, and LSCC patient-derived xenograft models. ERO1α is a functional downstream target of mTORC1. Elevated ERO1α induced ferroptosis resistance and exerted pro-oncogenic roles in mTORC1-activated cells via upregulation of SLC7A11. Mechanically, ERO1α stimulated the transcription of SLC7A11 by activating the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) pathway. Moreover, ERO1α inhibition combined with treatment using the ferroptosis inducer IKE exhibited synergistic antitumor effects on mTORC1-activated tumors. The ERO1α/IL-6/STAT3/SLC7A11 pathway is crucial for mTORC1-mediated ferroptosis resistance and tumor growth, and combining ERO1α inhibition with ferroptosis inducers is a novel and effective treatment for mTORC1-related tumors.

中文翻译：

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mTORC1 激活后增强的 ERO1α 通过上调 SLC7A11 诱导铁死亡抵抗和肿瘤进展

雷帕霉素复合物 1 (mTORC1) 信号传导失调的机制靶标在铁死亡抵抗和肿瘤发生中发挥着关键作用。然而，确切的潜在机制仍需要充分理解。通过实时定量 PCR (qRT-PCR)、蛋白质印迹、免疫荧光 (IF) 检测 mTORC1 激活的小鼠胚胎成纤维细胞、癌细胞和喉鳞状细胞癌 (LSCC) 临床样本中内质网氧化还原酶 1 α (ERO1α) 的表达）和免疫组织化学。我们进行了大量的体外和体内实验，以确定 ERO1α 及其下游靶标、溶质载体家族 7 的成员 11 (SLC7A11) 在 mTORC1 介导的细胞增殖、血管生成、铁死亡抵抗和肿瘤生长中的作用。通过 RNA 测序、细胞因子阵列、酶联免疫吸附测定、qRT-PCR、蛋白质印迹、IF、荧光素酶报告基因测定和染色质免疫沉淀测定研究 ERO1α 对 SLC7A11 的调节机制。使用细胞系来源的异种移植物、LSCC 类器官和 LSCC 患者来源的异种移植模型评估 ERO1α 抑制和铁死亡诱导剂咪唑酮erastin (IKE) 对 mTORC1 激活细胞的联合治疗效果。 ERO1α 是 mTORC1 的功能性下游靶标。 ERO1α 升高可诱导铁死亡抵抗，并通过上调 SLC7A11 在 mTORC1 激活细胞中发挥促癌作用。从机械角度来说，ERO1α 通过激活白细胞介素 6 (IL-6)/信号转导器和转录激活剂 3 (STAT3) 通路来刺激 SLC7A11 的转录。此外，ERO1α 抑制与铁死亡诱导剂 IKE 治疗相结合，对 mTORC1 激活的肿瘤表现出协同抗肿瘤作用。 ERO1α/IL-6/STAT3/SLC7A11 通路对于 mTORC1 介导的铁死亡抵抗和肿瘤生长至关重要，将 ERO1α 抑制与铁死亡诱导剂相结合是治疗 mTORC1 相关肿瘤的一种新颖且有效的治疗方法。