氧化应激诱导的生长抑制因子1（OSGIN1）调节细胞死亡。OSGIN1在非小细胞肺癌（NSCLC）中的作用和潜在的分子机制尚未被明确。我们使用免疫组织化学和免疫印迹法检测了NSCLC样本中OSGIN1的表达。通过MTT、软琼脂和成聚焦形成实验确定了表达OSGIN1或TUBB3敲下的NSCLC细胞和吉非替尼耐药细胞的生长情况。利用NSCLC患者衍生的异种移植瘤模型和吉非替尼耐药患者衍生的异种移植瘤模型评估了OSGIN1敲下对体内肿瘤生长的影响。利用免疫沉淀质谱/质谱分析、免疫沉淀、近扩增法和免疫印迹法鉴定了OSGIN1的潜在互作蛋白伴侣。通过微管聚合实验和免疫荧光分析探索了微管动力学。利用磷酸蛋白质组学、KEGG分析和免疫印迹法研究了OSGIN1敲下细胞中信号分子的差异表达。我们发现OSGIN1在NSCLC组织中高表达，并与肺癌患者低生存率和肿瘤大小呈正相关。OSGIN1敲下抑制了NSCLC细胞生长和体内患者衍生NSCLC肿瘤的生长。OSGIN1敲下强烈增加了微管的聚合，并在体外和体内恢复了对吉非替尼的敏感性。此外，TUBB3的敲下也强烈抑制了NSCLC细胞的增殖。在机制上，我们发现OSGIN1增强了DYRK1A介导的TUBB3丝氨酸172磷酸化，这对诱导微管解聚至关重要。磷酸蛋白质组学和本体分析结果表明OSGIN1敲下导致了MKK3/6-p38信号通路的传播减弱。我们提出OSGIN1通过增强DYRK1A介导的TUBB3丝氨酸172磷酸化来调节微管动力学。此外，高表达的OSGIN1通过MKK3/6-p38信号通路促进了NSCLC肿瘤的生长和吉非替尼耐药性。我们的发现揭示了OSGIN1的新机制，并为NSCLC临床治疗提供了一个有前景的治疗靶标。 © 2023年。作者，独家许可给Springer Nature Switzerland AG。

Oxidative stress induced growth inhibitor 1 (OSGIN1) regulates cell death. The role and underlying molecular mechanism of OSGIN1 in non-small cell lung cancer (NSCLC) are uncharacterized.OSGIN1 expression in NSCLC samples was detected using immunohistochemistry and Western blotting. Growth of NSCLC cells and gefitinib-resistant cells expressing OSGIN1 or TUBB3 knockdown was determined by MTT, soft agar, and foci formation assays. The effect of OSGIN1 knockdown on in vivo tumor growth was assessed using NSCLC patient-derived xenograft models and gefitinib-resistant patient-derived xenograft models. Potentially interacting protein partners of OSGIN1 were identified using IP-MS/MS, immunoprecipitation, PLA, and Western blotting assays. Microtubule dynamics were explored by tubulin polymerization assay and immunofluorescence. Differential expression of signaling molecules in OSGIN1 knockdown cells was investigated using phospho-proteomics, KEGG analysis, and Western blotting.We found that OSGIN1 is highly expressed in NSCLC tissues and is positively correlated with low survival rates and tumor size in lung cancer patients. OSGIN1 knockdown inhibited NSCLC cell growth and patient-derived NSCLC tumor growth in vivo. Knockdown of OSGIN1 strongly increased tubulin polymerization and re-established gefitinib sensitivity in vitro and in vivo. Additionally, knockdown of TUBB3 strongly inhibited NSCLC cell proliferation. Mechanistically, we found that OSGIN1 enhances DYRK1A-mediated TUBB3 phosphorylation, which is critical for inducing tubulin depolymerization. The results of phospho-proteomics and ontology analysis indicated that knockdown of OSGIN1 led to reduced propagation of the MKK3/6-p38 signaling axis.We propose that OSGIN1 modulates microtubule dynamics by enhancing DYRK1A-mediated phosphorylation of TUBB3 at serine 172. Moreover, elevated OSGIN1 expression promotes NSCLC tumor growth and gefitinib resistance through the MKK3/6-p38 signaling pathway. Our findings unveil a new mechanism of OSGIN1 and provide a promising therapeutic target for NSCLC treatment in the clinic.© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.