背景:最近的研究强调了NF-κB信号通路在癌症发生和发展中的重要作用。此外,长链非编码RNA(lncRNA)已被确定为维持NF-κB信号通路功能的关键调节因子。尽管有这些发现,lncRNAs通过其影响NF-κB途径的潜在分子机制仍未被研究。
方法:生物信息学分析用于研究XTP6的差异表达和预后意义。通过体外和体内实验方法进一步阐明了XTP6的功能作用。为了估计XTP6和NDH2之间的相互作用,进行RNA下拉和RNA免疫沉淀(RIP)测定。使用通过RNA纯化的染色质分离(ChIRP)测定检查XTP6和IκBα启动子之间的连接。此外,实施染色质免疫沉淀(ChIP)测定以分析c-myc与XTP6启动子的结合亲和力,提供对发挥作用的监管机制的见解。
结果:XTP6在多形性胶质母细胞瘤(GBM)组织中明显上调,并与GBM患者的不良预后有关。我们的研究表明,XTP6可以在体外和体内促进GBM的恶性进展。此外,XTP6通过募集NDH2到IκBα启动子下调IκBα表达,导致H3K27me3水平升高,从而降低了IκBα的转录活性。此外,c-myc介导的XTP6上调进一步推动了GBM的进展,并与IκBα建立了一个正反馈环,使NF-κB信号通路的激活得以延续.值得注意的是,靶向NF-κB信号通路的抑制剂的应用有效地抑制了XTP6诱导的持续激活,导致体内肿瘤形成的显著减少。
结论:结果表明,XTP6揭示了一种在NF-κB信号通路持续激活中发挥作用的创新表观遗传机制,为GBM的治疗提供了一个有希望的治疗靶点。
BACKGROUND: Recent studies have highlighted the significant role of the NF-κB signaling pathway in the initiation and progression of cancer. Furthermore, long noncoding RNAs (lncRNAs) have been identified as pivotal regulators in sustaining the NF-κB signaling pathway\'s functionality. Despite these findings, the underlying molecular mechanisms through which lncRNAs influence the NF-κB pathway remain largely unexplored.
METHODS: Bioinformatic analyses were utilized to investigate the differential expression and prognostic significance of XTP6. The functional roles of XTP6 were further elucidated through both in vitro and in vivo experimental approaches. To estimate the interaction between XTP6 and NDH2, RNA pulldown and RNA Immunoprecipitation (RIP) assays were conducted. The connection between XTP6 and the IκBα promoter was examined using Chromatin Isolation by RNA Purification (ChIRP) assays. Additionally, Chromatin Immunoprecipitation (ChIP) assays were implemented to analyze the binding affinity of c-myc to the XTP6 promoter, providing insights into the regulatory mechanisms at play.
RESULTS: XTP6 was remarkedly upregulated in glioblastoma multiforme (GBM) tissues and was connected with adverse prognosis in GBM patients. Our investigations revealed that XTP6 can facilitate the malignant progression of GBM both in vitro and in vivo. Additionally, XTP6 downregulated IκBα expression by recruiting NDH2 to the IκBα promoter, which resulted in elevated levels of H3K27me3, thereby reducing the transcriptional activity of IκBα. Moreover, the progression of GBM was further driven by the c-myc-mediated upregulation of XTP6, establishing a positive feedback loop with IκBα that perpetuated the activation of the NF-κB signaling pathway. Notably, the application of an inhibitor targeting the NF-κB signaling pathway effectively inhibited the continuous activation induced by XTP6, leading to a significant reduction in tumor formation in vivo.
CONCLUSIONS: The results reveal that XTP6 unveils an innovative epigenetic mechanism instrumental in the sustained activation of the NF-κB signaling pathway, suggesting a promising therapeutic target for the treatment of GBM.