Pomegranate for Chemoprevention and Chemotherapy of Prostate Cancer

描述（由申请人提供）：识别新靶点并开发用于预防和治疗前列腺癌（PCa）的常用无毒饮食制剂已成为国家医疗优先事项。最近的研究强调了转录因子NRcB在癌细胞存活中的作用。沿着这些路线，许多研究表明，NRcB在人前列腺癌细胞和标本中被组成型激活。因此，NRcB已成为开发新的PCa预防和治疗方法的重要靶点。从石榴树的果实中提取的石榴被认为具有抗氧化和抗炎的特性。我们最近发现，石榴果实提取物（PFE）通过抑制NF κ B信号通路抑制小鼠皮肤2阶段致癌模型中的肿瘤发展。我们未发表的初步数据表明，PFE是一种有效的抗增殖和促凋亡剂对人前列腺癌PCS细胞。通过MALDI-TOF质谱分析，发现PFE含有6种花青素和几种鞣花单宁和可水解单宁。我们的初步数据表明，在PFE中存在的6种花青素中，飞燕草素是最丰富的是最有效的抗增殖剂。重要的是，已知飞燕草素存在于许多其他色素水果和蔬菜中，如浆果，茄子，黑葡萄，番茄，胡萝卜和红洋葱。本提案的主要目的是利用我们在PCa细胞中使用飞燕草素和PFE的新的初步发现，并在体外和体内环境下研究其PCa化学预防和/或化学治疗潜力。之所以做出这样的选择，是因为我们认为细胞培养研究必须使用纯试剂而不是试剂的混合物进行，干预研究必须使用人类可以被说服消费的物质进行。使用PCa细胞系，我们将首先研究由飞燕草素激活的细胞死亡级联反应，并了解它如何抑制NF κ B介导的细胞存活信号。特别地，使用遗传操作和药理学方法，我们将研究NF κ B基因产物在i）抗凋亡（IAP 1、xIAP、Bfl-1/A1、Bcl-2、cFLIP和存活素），ii）增殖（细胞周期蛋白D1和c-Myc），iii）炎症（考克斯-2、PPARgamma和iNOS）和iv）转移（MMP-9、MMP-2和VEGF）中的作用。然后，我们将使用已建立的转基因和异种移植小鼠模型在体内情况下定义PFE的PCa化学预防/治疗潜力。为了与人类群体相关，我们推断疗效研究必须在疾病进展发生类似于人类的动物模型中进行。采用i）TRAMP，一种模拟人类前列腺疾病的进行性形式的模型和ii）植入人类PCa细胞的无胸腺裸鼠，我们将建立PFE的癌症化学预防/治疗潜力。公众相关性：该提案将建立前列腺癌预防和治疗的分子靶点，此外，还将提请注意使用石榴和其他水果和蔬菜预防前列腺癌。

DESCRIPTION (provided by applicant): Identification of novel targets and the development of commonly used and non-toxic dietary agents for prevention and treatment of prostate cancer (PCa) has become a National Medical Priority. Recent studies have highlighted the role of the transcription factor NRcB in cancer cell survival. Along these lines, many studies have shown that NRcB is constitutively activated in human PCa cells and specimens. Thus, NRcB has become an important target for developing novel PCa preventive and therapeutic approaches. Pomegranate derived from the fruit of the tree Punica granatum is known to possess antioxidant and anti-inflammatory properties. We recently showed that pomegranate fruit extract (PFE) inhibited tumor development in mouse skin 2 stage carcinogenesis model through inhibition of the NFkappaB signaling pathway. Our unpublished preliminary data showed that PFE is an effective anti-proliferative and pro-apoptotic agent against human PCa PCS cells. On analysis by MALDI-TOF mass spectrometry, PFE was found to contain 6 anthocyanins and several ellagitannins and hydrolyzable tannins. Our preliminary data showed that among 6 anthocyanins present in PFE, delphinidin the most abundant was the most effective anti-proliferative agent. Importantly, delphinidin is known to be present in many other pigmented fruits and vegetables like berries, egg plant, dark grapes, tomato, carrots and red onions. The primary objective of this proposal is to capitalize on our novel preliminary findings with delphinidin and PFE in PCa cells and investigate its PCa chemopreventive and/or chemotherapeutic potential under in vitro and in vivo settings. This choice is made because we believe that cell culture studies must be conducted with pure agents rather than a mixture of agents and intervention studies must be conducted with the substances that humans could be persuaded to consume. Using PCa cell lines, we will first investigate the cell death cascade activated by delphinidin and understand how it inhibits cell survival signaling mediated by NFkappaB. In particular, using genetic manipulation and pharmacological approaches, we will investigate the role of NFkappaB gene products involved in i) anti-apoptosis (IAP1, xlAP, Bfl-1/A1, Bcl-2, cFLIP, and survivin), ii) proliferation (cyclin D1 and c-Myc), iii) inflammation (Cox-2, PPARgamma and iNOS) and iv) metastasis (MMP-9, MMP-2 and VEGF). We will then define PCa chemopreventive/therapeutic potential of PFE under in vivo situation using established transgenic and xenograft mouse models. To have relevance to human population, we reasoned that efficacy studies must be conducted in an animal model where disease progression occurs akin to humans. Employing i) TRAMP, a model that mimics progressive forms of human prostatic disease and ii) athymic nude mouse implanted with human PCa cells, we will establish the cancer chemopreventive/therapeutic potential of PFE. PUBLIC RELEVANCE: This proposal will establish the molecular targets for prostate cancer prevention and treatment and in addition, will draw attention to the use of pomegranate and other piamented fruits and vegetables for prevention against prostate cancer.