The NF-κB Transcription Factor Pathway as a Therapeutic Target in Cancer: Methods for Detection of NF-κB Activity
The NF-κB Transcription Factor Pathway as a Therapeutic Target in Cancer: Methods for Detection of NF-κB Activity
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DOI：
10.1007/978-1-60327-530-9_10
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发表时间：
2009-01-01
期刊：
影响因子：
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通讯作者：
Franzoso, Guido
中科院分区：
文献类型：
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作者：
Mauro, Claudio;Zazzeroni, Francesca;Franzoso, Guido
NF-κ B转录因子调控先天性和适应性免疫和炎症。NF-κ B还对抗由促炎细胞因子肿瘤坏死因子（TNF）α诱导的程序性细胞死亡（PCD），并且NF-κ B的这种活性对于生物体生理学、慢性炎症和肿瘤发生至关重要。事实上，尽管NF-κ B对肿瘤发生的许多方面都有贡献，但现在清楚的是，它对PCD的抑制作用是该过程的核心。值得注意的是，最近的研究表明，NF-κ B B代表了炎症和癌变之间的明确关联的关键环节。在这一联系中，NF-κ B促进刺激癌细胞生长的炎性介质（例如TNF α）的合成，并上调保护这些细胞免受炎性信号诱导的PCD的基因。升高的NF-κ B活性也阻碍了放射和化疗药物对肿瘤细胞的杀伤，从而促进了对抗癌治疗的抵抗。因此，NF-κ B靶向药物越来越多地用于治疗人类恶性肿瘤。然而，由于NF-κ B途径的普遍存在的性质，这些药物具有严重的副作用，这限制了它们的临床应用。因此，优选的方法是阻断NF-κ B的关键下游靶点，而不是阻断NF-κ B本身，所述靶点介导癌症中的离散功能，例如促生存功能。最近的发现揭示了用于控制PCD的NF-κ B诱导机制中的组织特异性，并鉴定了这种控制的推定效应物，这清楚地验证了这种治疗方法。考虑到肿瘤坏死因子α诱导的NF-κ B活化信号在癌症中的作用以及这些信号产生新的抗癌疗法的潜力，我们在此集中于最常用于分析从触发TNF-受体（TNF-R）1（TNF α的主要受体）到诱导NF-κ B的分子步骤的方法。具体而言，我们综述了用于分析TNF-R1运输、所谓的TNF-R1复合物1的组装、I κ B激酶（IKK）复合物的形成和激活、抑制性I κ B蛋白的磷酸化和蛋白水解、NF-κ B二聚体的翻译后修饰和核转位、NF-κ B转录活性的诱导以及与特异性启动子的结合的方法，和NF-κ B靶基因的上调。对癌细胞中这些事件的分析不仅可以更好地理解NF-κ B B在癌发生中的作用，而且可以为选择性抗癌治疗提供潜在的新靶点。
NF-kappa B transcription factors marshal innate and adaptive immunity and inflammation. NF-kappa B also counters programmed cell death (PCD) induced by the proinflammatory cytokine tumor necrosis factor (TNF)alpha, and this activity of NF-kappa B is crucial for organismal physiology, chronic inflammation, and tumorigenesis. Indeed, whereas NF-kappa B contributes to many aspects of oncogenesis, it is now clear that its suppressive action on PCD is central to this process. Notably, recent studies indicate that NF-kappa B represents a crucial link in the well-established association between inflammation and carcinogenesis. In this link, NF-kappa B promotes synthesis of inflammatory mediators (e.g. TNF alpha) that stimulate growth of cancer cells, and upregulates genes that protect these cells against PCD induced by inflammatory signals. Elevated NF-kappa B activity also hampers tumor-cell killing inflicted by radiation and chemotherapeutic drugs, and in so doing, promotes resistance to anticancer therapy. Accordingly, NF-kappa B-targeting drugs are increasingly being used for treatment of human malignancies. Owing to the ubiquitous nature of the NF-kappa B pathway, however, these drugs have serious side effects, which limit their clinical use. Thus, a preferable approach would be to block, rather than NF-kappa B itself, its critical downstream targets that mediate discrete functions in cancer, such as prosurvival functions. Recent discoveries unraveling tissue specificity in the NF-kappa B-inducible mechanism(s) for control of PCD and identifying putative effectors of this control clearly validate this therapeutic approach. Given the emerging role of TNF alpha-induced signals of NF-kappa B activation in cancer and the potential of these signals for yielding new anticancer therapies, we focus herein on the methods most commonly used for analysis of the molecular steps leading from the triggering of TNF-Receptor (TNF-R)1 - the primary receptor of TNF alpha - to the induction of NF-kappa B. Specifically, we review the methods used for analysis of TNF-R1 trafficking, assembly of so-called TNF-R1 complex 1, formation and activation of the I kappa B kinase (IKK) complex, phosphorylation and proteolysis of inhibitory I kappa B proteins, post-translational modifications and nuclear translocation of NF-kappa B dimers, induction of NF-kappa B transcriptional activity and binding to specific promoters, and upregulation of NF-kappa B target genes. The analysis of these events in cancerous cells may not only provide a better understanding of the basis for the role of NF-kappa B in carcinogenesis, but also potential new targets for selective anticancer therapy.