Background/Objectives: RAF pathway aberrations are one of the hallmarks of lung cancer. Sorafenib is a multi-kinase inhibitor targeting the RAF pathway and is FDA-approved for several cancers, yet its efficacy in lung cancer is controversial. Previous clinical research showed that aARAFp.S214C mutation exhibited exceptional responsiveness to sorafenib in lung adenocarcinoma.Methods: Considering this promising clinical potential, the oncogenic potential and sorafenib response of theARAFp.S214C mutation were investigated using lung cancer models.ARAFp.S214C mutant,ARAFwild-type (WT), andEGFPcontrol genes were ectopically expressed in lung adenocarcinoma cell lines retroviral transduction. In vitro and in vivo sorafenib sensitivity studies were performed, followed by transcriptomics and proteomics analyses.Results: Compared to theARAF-WT andEGFP-engineered cells, theARAFp.S214C-engineered cells activated Raf-MEK-ERK signaling and exhibited enhanced oncogenic potential in terms of in vitro cell proliferation, colony and spheroid formation, migration, and invasion abilities, as well as in vivo tumorigenicity. TheARAFp.S214C-engineered cells also displayed heightened sensitivity to sorafenib in vitro and in vivo. RNA sequencing and reverse-phase protein array analyses demonstrated elevated expression of genes and proteins associated with tumor aggressiveness in theARAFp.S214C mutants, and its sorafenib sensitivity was likely moderated through inhibition of the cell cycle and DNA replication. The ERK and PI3K signaling pathways were also significantly deregulated in theARAFp.S214C mutants regardless of sorafenib treatment.Conclusions: This study demonstrates the oncogenicity and sorafenib sensitivity of theARAFp.S214C mutation in lung cancer cells, which may serve as a biomarker for predicting the sorafenib response in lung cancer patients. Importantly, investigating the gene–drug sensitivity pairs in clinically exceptional responders may guide and accelerate personalized cancer therapies based on specific tumor mutations.
背景/目的：RAF通路异常是肺癌的标志性特征之一。索拉非尼是一种靶向RAF通路的多激酶抑制剂，已获美国FDA批准用于多种癌症治疗，但其在肺癌中的疗效尚存争议。先前临床研究表明，ARAF p.S214C突变在肺腺癌中对索拉非尼表现出显著的治疗反应。 方法：基于这一具有前景的临床潜力，本研究利用肺癌模型探究ARAF p.S214C突变的致癌潜力及对索拉非尼的治疗反应。通过逆转录病毒转导技术，在肺腺癌细胞系中外源性表达ARAF p.S214C突变体、ARAF野生型（WT）及EGFP对照基因。随后开展体外和体内索拉非尼敏感性研究，并进行转录组学与蛋白质组学分析。 结果：与ARAF-WT和EGFP工程化细胞相比，ARAF p.S214C工程化细胞能激活Raf-MEK-ERK信号通路，并在体外细胞增殖、克隆形成、球体形成、迁移和侵袭能力，以及体内致瘤性方面表现出更强的致癌潜力。同时，ARAF p.S214C工程化细胞在体外和体内对索拉非尼均显示出更高的敏感性。RNA测序和反相蛋白芯片分析表明，ARAF p.S214C突变体中与肿瘤侵袭性相关的基因和蛋白表达上调，其索拉非尼敏感性可能通过抑制细胞周期和DNA复制来调节。无论是否接受索拉非尼治疗，ARAF p.S214C突变体中的ERK和PI3K信号通路均出现显著失调。 结论：本研究证实了ARAF p.S214C突变在肺癌细胞中的致癌性及对索拉非尼的敏感性，该突变或可作为预测肺癌患者索拉非尼治疗反应的生物标志物。重要的是，对临床特殊应答者中基因-药物敏感性配对的研究，可基于特定肿瘤突变为指导和加速个体化癌症治疗提供依据。
Oncogenic Activity and Sorafenib Sensitivity ofARAFp.S214C Mutation in Lung Cancer
肿瘤(癌症)患者之家