Producing 14C-Phytoene & Phytofluene for Cancer Research

描述（由申请人提供）：前列腺癌是美国男性癌症死亡的第二大原因。流行病学证据表明，增加番茄制品的消费量可以降低患前列腺癌的风险。西红柿含有多种类胡萝卜素，其中一些已被证明具有抗氧化和抗癌作用。最近的研究主要集中在番茄红素这一初级类胡萝卜素上，但对番茄红素、植物烯和植物荧光素的类胡萝卜素生物合成前体的研究很少。我们假设，除了番茄红素，植物烯和植物流感素有助于降低前列腺癌的风险。目前尚不清楚植物烯和植物荧光素在前列腺中的吸收、代谢和可用性如何，因为植物烯和植物荧光素没有商业上的可用性。必须开发可靠的方法来生产、分离和分析这些类胡萝卜素。这项工作的总体目标是优化番茄细胞悬浮培养系统，以生物合成14c放射性标记的植物烯和植物荧光素，以研究这些类胡萝卜素在动物模型中的生物利用度、代谢和生物分布。本研究的具体目的是：1)利用体外番茄细胞悬浮培养系统，通过添加植物烯去饱和酶抑制剂去氟拉松产生14c -放射性标记的植物烯和植物流感素；2)优化从番茄细胞悬浮培养中提取和分离14c -放射性标记的植物烯和植物流感素；3)确定14c -放射性标记的植物烯和植物流感素在F344大鼠模型中的生物利用度、代谢和生物分布。这项工作的结果将是开发新的和可靠的方法来生产和分离14c放射性标记的植物烯和植物荧光素，以及确定植物烯、植物荧光素及其代谢物在前列腺中的积累。这些新方法将提供在动物模型中设计体内癌症预防试验和开发用于人体代谢工作的13c标记类胡萝卜素的能力。

DESCRIPTION (provided by applicant): Prostate cancer is the second leading cause of cancer death in American men. Epidemiological evidence indicates a decreased risk of prostate cancer with increased consumption of tomato products. Tomatoes contain a variety of carotenoids, some of which have been shown to have antioxidant and anticancer effects. Recent research has focused on the primary tomato carotenoid, lycopene, yet few studies have been conducted with the carotenoid biosynthetic precursors of lycopene, phytoene and phytofluene. We hypothesize that in addition to lycopene, phytoene and phytofluene contribute to the reduced risk of prostate cancer. It is unclear how well phytoene and phytofluene are absorbed, metabolized, and made available for use in the prostate, because phytoene and phytofluene are not commercially available. Reliable methods to produce, isolate, and analyze these carotenoids must be developed. The overall objective of this work is to optimize a tomato cell suspension culture system to biosynthesize 14C-radiolabeled phytoene and phytofluene for bioavailability, metabolism, and biodistribution studies of these carotenoids in an animal model. The specific aims of this proposal are: 1) To utilize an in vitro tomato cell suspension culture system to produce both 14C-radiolabeled phytoene and phytofluene through the addition of a phytoene desaturase inhibitor, norflurazon, 2) To optimize the extraction and separation of 14C-radiolabeled phytoene and phytofluene from tomato cell suspension cultures, 3) To determine the bioavailability, metabolism, and biodistribution of 14C-radiolabeled phytoene and phytofluene in the F344, rat model. The outcome of this work will be the development of novel and reliable methods to produce and isolate 14C-radiolabeled phytoene and phytofluene, in addition to determining the accumulation of phytoene, phytofluene, and their metabolites in the prostate. These novel methods will provide the ability to design both an in vivo cancer prevention trial in an animal model and to develop 13C-labeled carotenoids for human metabolic work.