Neeman M, Degani H
Isotope Department, Weizmann Institute of Science, Rehovot, Israel.
Proc Natl Acad Sci U S A. 1989 Jul;86(14):5585-9. doi: 10.1073/pnas.86.14.5585.
Metabolic changes following estrogen stimulation and the inhibition of these changes in the presence of actinomycin D and cycloheximide were monitored continuously in perfused human breast cancer T47D clone 11 cells with 31P and 13C NMR techniques. The experiments were performed by estrogen rescue of tamoxifen-treated cells. Immediately after perfusion with estrogen-containing medium, a continuous enhancement in the rates of glucose consumption, lactate production by glycolysis, and glutamate synthesis by the Krebs cycle occurred with a persistent 2-fold increase at 4 hr. The content of phosphocholine had increased by 10% to 30% within the first hour of estrogen stimulation, but the content of the other observed phosphate metabolites as well as the pH remained unchanged. Pretreatment with either actinomycin D or cycloheximide, at concentrations known to inhibit mRNA and protein synthesis, respectively, and simultaneous treatment with estrogen and each inhibitor prevented the estrogen-induced changes in glucose metabolism. This suggested that the observed estrogen stimulation required synthesis of mRNA and protein. These inhibitors also modulated several metabolic activities that were not related to estrogen stimulation. The observed changes in the in vivo kinetics of glucose metabolism may provide a means for the early detection of the response of human breast cancer cells to estrogen versus tamoxifen treatment.
采用³¹P和¹³C NMR技术，对灌注的人乳腺癌T47D克隆11细胞中雌激素刺激后的代谢变化以及在放线菌素D和环己酰亚胺存在下这些变化的抑制情况进行了连续监测。实验通过对他莫昔芬处理的细胞进行雌激素挽救来进行。在用含雌激素的培养基灌注后，葡萄糖消耗率、糖酵解产生乳酸的速率以及三羧酸循环合成谷氨酸的速率立即持续增强，在4小时时持续增加两倍。在雌激素刺激的第一小时内，磷酸胆碱含量增加了10%至30%，但其他观察到的磷酸盐代谢物含量以及pH值保持不变。分别用已知可抑制mRNA和蛋白质合成的浓度的放线菌素D或环己酰亚胺进行预处理，以及同时用雌激素和每种抑制剂进行处理，可防止雌激素诱导的葡萄糖代谢变化。这表明观察到的雌激素刺激需要mRNA和蛋白质的合成。这些抑制剂还调节了几种与雌激素刺激无关的代谢活动。观察到的体内葡萄糖代谢动力学变化可能为早期检测人乳腺癌细胞对雌激素与他莫昔芬治疗的反应提供一种方法。
