ESTROGEN-INDUCED CHANGES IN BREAST CANCER CELLS

这项研究的长期目的是开发一种非 MRS介入性方法用于雌激素的表征 乳腺癌的反应性。为此，具体而言， 雌激素和抗雌激素调节的代谢活动 将在人类乳腺癌细胞系中表现为 类固醇激素受体：MCF-7、T47D和ZR-75-1。 这项研究是我们总体努力的一部分 荷尔蒙引起生殖器官的变化和 相关恶性肿瘤(见附录)。 搜索将集中在已知的活动上 雌激素在真正的靶器官中的增强作用，例如：1) 肌酸激酶(BB同工酶)的活性，这是一种受欢迎的早期 雌激素作用的标志物；2)雌激素作用的几个关键酶 糖酵解途径；3)葡萄糖-6-磷酸脱氢酶 (G6PD)，为其增加了mRNA的生产 观察到的。此外，预计监控的能力 在活跃的生长细胞中的在线新陈代谢将导致 发现不可预测的变化，就像 磷脂酰胆碱和甘油衍生物的浓度 磷酰乙醇胺。 核磁共振波谱将是这项研究的主要工具。方法 在核磁共振过程中保持细胞的存活、活跃和生长 开发了测量方法。将采用31-P核磁共振技术 测定细胞的能量学和动力学 磷酸盐转移反应。13-C核磁共振研究将主要 专注于标记底物的实时跟踪， 中间体和提供熔剂量度的终端产品 通过糖酵解酶和G6PD。1-氢核磁共振 测量和1-H观测到的13-C去耦合序列将 补充13-C结果。方法和结果 这项研究的结果将随后用于这些的MRS研究 人乳腺癌细胞系，移植到裸鼠体内，以及 人类的乳房肿瘤。

The long range purpose of this research is to develop a non invasive method using MRS for the characterization of estrogen responsiveness of breast carcinoma. To that end, specific metabolic activities modulated by estrogens and by antiestrogens will be characterized in human breast cancer cell lines that retain the receptors for steroid hormones: MCF-7, T47D and ZR-75-1. This research is part of our general efforts to characterize hormonal induced changes in the reproductive organs and associated malignancies (see Appendix). The search will concentrate on activities that are known to be enhanced by estrogen in genuine target organs such as: 1) the activity of creatine kinase (the BB isozyme), a favored early marker of estrogen action; 2) several key enzymes of the glycolytic pathway; and 3) glucose-6-phosphate dehydrogenase (G6PD), for which increased production of mRNA has been observed. In addition, it is anticipated that the ability to monitor the metabolism on line, in active growing cells, will lead to the discovery of unpredicted changes as seem to occur with the concentration of the glycerol derivatives of phosphorylcholine and phosphorylethanolamine. NMR spectroscopy will be the main tool in this study. Methods for keeping the cells alive, active and growing during the NMR measurements were developed. 31-P NMR will be employed for determining the energetics of the cells and the kinetics of phosphate transfer reactions. 13-C NMR studies will mainly concentrate on real time follow up of labeled substrates, intermediates, and end products providing a measure of the fluxes through the glycolytic enzymes and G6PD. 1-H NMR measurements and 1-H observed 13-C decoupled sequences will complement the 13-C results. The methodologies and the results of this research will subsequently be used in MRS studies of these human breast cancer cell lines, implanted in nude mice, and of breast tumors in humans.