Role of E2F1 in eugenol-mediated antiproliferative activity in melanoma cells

描述（由申请人提供）： 晚期转移性黑色素瘤患者尚无有效的标准治疗。我们已经证明丁香酚是一种体外和体内抗黑色素瘤药物。该探索性提案的目的是建立以下原则的证据：（i）丁香酚的最安全和最有效剂量将取决于其分配的途径，以及（ii）E2 F1是黑色素瘤细胞中的丁香酚靶点。我们的工作假设是，在黑色素瘤细胞中，丁香酚诱导的DNA损伤存在双相反应。我们已经提出，丁香酚处理后，DNA损伤信号通过ATM/chk 2途径发生，导致E2 F1的稳定。此外，作为DNA损伤的函数，存在E2 F1对丁香酚的响应的时间分离。E2 F1作为一种即时反应，反式激活ogg 1以修复由丁香酚引起的DNA碱基损伤。然而，随着DNA损伤持续，E2 F1转录活性被抑制。作为PCNA表达的结果，不出现E2 F1靶基因，因此细胞被阻滞在S期。我们还将确定使用1205 Lu异种移植模型递送丁香酚的最有效方法。在这个目标中，我们还将确定最安全（无毒）的丁香酚剂量。这项探索性建议的结果将决定丁香酚是否可以进一步开发为抗黑色素瘤药物。这一提议的意义在于，丁香酚靶向黑色素瘤中的失调途径，并且因为E2 F1调节参与细胞过程（如凋亡和转移）的几个基因，丁香酚可以影响这些途径。目前还没有治疗晚期黑色素瘤的药物。本研究目标的完成将导致后续提案（RO 1），旨在更好地了解丁香酚的作用机制及其在黑色素瘤临床前模型中的临床效用。公共卫生相关性：由于转移性恶性黑色素瘤缺乏有效的治疗选择，死亡率继续攀升。本提案的目的是确定给予该化合物的最有效方式，并了解其生长抑制活性的机制。该项目的完成将使我们更接近于制定预防致命性转移性黑色素瘤的策略。

DESCRIPTION (provided by applicant): There is no effective standard therapy for patients with advanced metastatic melanoma. We have shown that eugenol, is an anti-melanoma agent in vitro and in vivo. The aim of this exploratory proposal is to establish proof of principle that (i) the safest and most effective dose of eugenol will depend on the route through which it is dispensed and (ii) E2F1 is a eugenol target in melanoma cells. Our working hypothesis is that there is a biphasic response to eugenol-induced DNA damage in melanoma cells. We have proposed that following eugenol treatment DNA damage signaling occurs through ATM/chk2 pathway that leads to stabilization of E2F1. Further there is temporal separation of E2F1 responses to eugenol as a function of DNA damage. As an immediate response E2F1 transactivates ogg1 to repair DNA base damage caused by eugenol. However as DNA damage persists E2F1 transcription activity is inhibited. As a result expression of PCNA, an E2F1 target gene does not occur and consequently cells are arrested in S phase. We will also determine the most efficacious method to deliver eugenol using the 1205Lu xenograft model. In this aim we will also determine the safest (non-toxic) eugenol dose. Results of this exploratory proposal will determine whether eugenol can be developed further as an anti-melanoma agent. Significance of this proposal is that eugenol targets a deregulated pathway in melanoma and because E2F1 regulates several genes involved in cellular processes such as apoptosis and metastasis eugenol can impact these pathways. There are no therapeutic agents in the immediate pipeline for advanced melanoma. Completion of the goals of this study will lead to subsequent proposals (RO1) aimed at obtaining a better understanding of the mechanism of action of eugenol and its clinical utility in preclinical models of melanoma. PUBLIC HEALTH RELEVANCE: Due to the lack of effective treatment options for metastatic malignant melanoma death rate continues to climb. The goal of the current proposal is to determine the most effective way to administer the compound and understand the mechanism of its growth inhibition activity. Completion of this project will take us closer to developing strategies to prevent fatal metastatic melanoma.