Targeted therapy and immunotherapy have led to dramatic change in the treatment of lung cancer, however, the overall 5-year survival rate of lung cancer patients is still suboptimal. It is important to exploit new potential of molecularly targeted therapies. High-frequency somatic mutations in KEAP1/NRF2 (27.9%) have been identified in lung squamous cell carcinoma. In this research, we explored the role of KEAP1 somatic mutations in the development of LSCC and whether a nuclear factor erythroid 2-related factor 2(NRF2) inhibitor be potential to target lung cancer carrying KEAP1/NRF2 mutations. Lung cancer cell lines A549 and H460 with loss-of-function mutations in KEAP1 stably transfected with wild-type (WT) KEAP1 or somatic mutations in KEAP1 were used to investigate the functions of somatic mutations in KEAP1. Flow cytometry, plate clone formation experiments, and scratch tests were used to examine reactive oxygen species, proliferation, and migration of these cell lines. The expression of NRF2 and its target genes increased, and tumor cell proliferation, migration, and tumor growth were accelerated in A549 and H460 cells stably transfected with KEAP1 mutants compared to control cells with a loss-of-function KEAP1 mutation and stably transfected with WT KEAP1 in both in vitro and in vivo studies. The proliferation of A549 cell line trasfected with the R320Q KEAP1 mutant was inhibited more apparent than that of the A549 cell line trasfected with WT KEAP1 after treatment with NRF2 inhibitor ML385. Somatic mutations of KEAP1 identified from patients with LSCC likely promote tumorigenesis mediated by activation of the KEAP1/NRF2 antioxidant stress response pathway. NRF2 inhibition with ML385 could inhibit the proliferation of tumor cells with KEAP1 mutation.

中文翻译：

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KEAP1 功能丧失突变通过 KEAP1/NRF2 通路激活驱动肺癌进展。

靶向治疗和免疫治疗使肺癌的治疗发生了巨大的变化，但肺癌患者的整体5年生存率仍不理想。开发分子靶向治疗的新潜力很重要。KEAP1/NRF2 中的高频体细胞突变（27.9%）已在肺鳞状细胞癌中被发现。在这项研究中，我们探讨了 KEAP1 体细胞突变在 LSCC 发展中的作用，以及核因子红细胞 2 相关因子 2（NRF2）抑制剂是否有潜力靶向携带 KEAP1/NRF2 突变的肺癌。使用野生型(WT) KEAP1 或KEAP1 体细胞突变稳定转染的KEAP1 功能丧失突变的肺癌细胞系A549 和H460 来研究KEAP1 体细胞突变的功能。流式细胞术，平板克隆形成实验和划痕测试用于检查这些细胞系的活性氧、增殖和迁移。与功能缺失的KEAP1突变并稳定转染WT的对照细胞相比，稳定转染KEAP1突变体的A549和H460细胞中NRF2及其靶基因的表达增加，肿瘤细胞增殖、迁移和肿瘤生长加速KEAP1 在体外和体内研究中。在用 NRF2 抑制剂 ML385 处理后，用 R320Q KEAP1 突变体转染的 A549 细胞系的增殖比用 WT KEAP1 转染的 A549 细胞系的增殖受到更明显的抑制。从 LSCC 患者中鉴定出的 KEAP1 体细胞突变可能促进了由 KEAP1/NRF2 抗氧化应激反应通​​路激活介导的肿瘤发生。