Campbell Jessica K, Rogers Randy B, Lila Mary Ann, Erdman John W
Division of Nutritional Sciences, University of Illinois at Urbana--Champaign, 905 South Goodwin Avenue, Illinois 61801, USA.
J Agric Food Chem. 2006 Feb 8;54(3):747-55. doi: 10.1021/jf0581269.
This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.
这项工作描述了一种植物细胞培养生产方法的开发和应用，用于生物合成和放射性标记八氢番茄红素和六氢番茄红素，以进行前列腺癌细胞培养研究。将除草剂氟草敏添加到已建立的番茄（Lycopersicon esculentum cv. VFNT樱桃）细胞悬浮培养物中，以诱导番茄红素前体八氢番茄红素和六氢番茄红素以其天然异构体形式（15-顺式八氢番茄红素和两种顺式六氢番茄红素异构体）的生物合成和积累。对氟草敏浓度、溶剂载体类型和浓度以及培养物暴露于氟草敏的持续时间进行了筛选，以优化八氢番茄红素和六氢番茄红素的合成。在用0.07%溶剂载体提供的0.03 mg氟草敏/40 mL新鲜培养基处理7天后，八氢番茄红素和六氢番茄红素均达到最大产量。向番茄细胞培养基中引入14C-蔗糖能够生产14C标记的八氢番茄红素，用于后续前列腺肿瘤细胞摄取研究。在DU 145前列腺肿瘤细胞中，确定15-顺式八氢番茄红素和八氢番茄红素的一种氧化产物被细胞摄取并部分代谢。从番茄细胞培养物中生物合成、放射性标记和分离这些类胡萝卜素的能力是一种新颖、有价值的方法，可用于进一步体外和体内研究八氢番茄红素和六氢番茄红素在癌症化学预防中的作用。
