背景:新的证据表明N6-甲基腺苷(m6A)RNA修饰在肿瘤发生和肿瘤进展中的关键作用。然而,m6A在非小细胞肺癌(NSCLC)中的作用尚不清楚.本研究旨在探讨m6A去甲基酶脂肪量和肥胖相关蛋白(FTO)在NSCLC肿瘤转移中的作用。
方法:人m6A表位基因组微阵列分析用于鉴定FTO的下游靶标。定量实时PCR(qRT-PCR)和蛋白质印迹用于评估FTO和FAP在NSCLC细胞系和组织中的表达水平。在体内和体外进行功能增益和功能丧失测定以评估FTO和FAP对NSCLC转移的影响。M6A-RNA免疫沉淀(MeRIP),RNA免疫沉淀(RIP),荧光素酶报告基因测定,和RNA稳定性分析用于探索FTO作用机制。免疫共沉淀(co-IP)测定用于确定FAP在NSCLC转移中的机制。
结果:FTO上调,预测NSCLC患者预后不良。FTO促进非小细胞肺癌细胞迁移和侵袭,FAK抑制剂defactinib(VS6063)抑制FTO过表达诱导的NSCLC转移。机械上,FTO通过以YTHDF2依赖性方式改变FAP的m6A水平促进NSCLC转移。此外,FTO介导的转移形成依赖于FAP和整合素家族成员之间的相互作用,这进一步激活了FAK信令。
结论:我们目前的发现为FTO介导的m6A去甲基化修饰在NSCLC转移中的作用提供了有价值的见解。通过激活FAP/整合素/FAK信号,FTO被确定为NSCLC转移的贡献者。这可能是NSCLC的潜在治疗靶点。视频摘要。
新的证据表明N6-甲基腺苷(m6A)RNA修饰在肿瘤发生和发展中的关键作用。尽管如此,m6A在NSCLC中的作用尚不清楚.目的探讨m6A去甲基酶脂肪量和肥胖相关蛋白(FTO)在非小细胞肺癌(NSCLC)肿瘤转移中的作用。结果表明,FTO上调,预测NSCLC患者预后不良。FTO促进非小细胞肺癌细胞迁移和侵袭,FAK抑制剂defactinib(VS6063)抑制FTO过表达诱导的NSCLC转移。机械上,FTO通过以YTHDF2依赖性方式改变FAP的m6A水平促进NSCLC转移。此外,FTO介导的转移形成依赖于FAP和整合素家族成员之间的相互作用,这进一步激活了FAK信令。我们目前的发现为FTO介导的m6A去甲基化修饰在NSCLC转移中的作用提供了有价值的见解。通过激活FAP/整合素/FAK信号,FTO被确定为NSCLC转移的贡献者。这可能是NSCLC的潜在治疗靶点。
Emerging evidence suggests the critical roles of N6-methyladenosine (m6A) RNA modification in tumorigenesis and tumor progression. However, the role of m6A in non-small cell lung cancer (NSCLC) is still unclear. This study aimed to explore the role of the m6A demethylase fat mass and obesity-associated protein (FTO) in the tumor metastasis of NSCLC.
A human m6A epitranscriptomic microarray analysis was used to identify downstream targets of FTO. Quantitative real-time PCR (qRT‒PCR) and western blotting were employed to evaluate the expression levels of FTO and FAP in NSCLC cell lines and tissues. Gain-of-function and loss-of-function assays were conducted in vivo and in vitro to assess the effects of FTO and FAP on NSCLC metastasis. M6A-RNA immunoprecipitation (MeRIP), RNA immunoprecipitation (RIP), luciferase reporter assays, and RNA stability assays were used to explore the mechanism of FTO action. Co-immunoprecipitation (co-IP) assays were used to determine the mechanism of FAP in NSCLC metastasis.
FTO was upregulated and predicted poor prognosis in patients with NSCLC. FTO promoted cell migration and invasion in NSCLC, and the FAK inhibitor defactinib (VS6063) suppressed NSCLC metastasis induced by overexpression of FTO. Mechanistically, FTO facilitated NSCLC metastasis by modifying the m6A level of FAP in a YTHDF2-dependent manner. Moreover, FTO-mediated metastasis formation depended on the interactions between FAP and integrin family members, which further activated the FAK signaling.
Our current findings provided valuable insights into the role of FTO-mediated m6A demethylation modification in NSCLC metastasis. FTO was identified as a contributor to NSCLC metastasis through the activation of the FAP/integrin/FAK signaling, which may be a potential therapeutic target for NSCLC. Video Abstract.
Emerging evidence suggests the crucial roles of N6-methyladenosine (m6A) RNA modification in tumorigenesis and progression. Nonetheless, the role of m6A in NSCLC remains unclear. The purpose of this study was to investigate the role of m6A demethylase fat mass and obesity-associated protein (FTO) in the tumor metastasis of non-small cell lung cancer (NSCLC). Results illustrated that FTO was upregulated and predicted poor prognosis in NSCLC patients. FTO promoted cell migration and invasion in NSCLC, and the FAK inhibitor defactinib (VS6063) suppressed NSCLC metastasis induced by overexpression of FTO. Mechanistically, FTO facilitated NSCLC metastasis by modifying the m6A level of FAP in a YTHDF2-dependent manner. Moreover, FTO-mediated metastasis formation depended on the interactions between FAP and integrin family members, which further activated the FAK signaling. Our current findings provided valuable insights into the role of FTO-mediated m6A demethylation modification in NSCLC metastasis. FTO was identified as a contributor to NSCLC metastasis through the activation of the FAP/integrin/FAK signaling, which may be a potential therapeutic target for NSCLC.