Lipid metabolism in T47D human breast cancer cells: 31P and 13C-NMR studies of choline and ethanolamine uptake.
Lipid metabolism in T47D human breast cancer cells: 31P and 13C-NMR studies of choline and ethanolamine uptake.
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T47D 人乳腺癌细胞中的脂质代谢：胆碱和乙醇胺摄取的 31P 和 13C-NMR 研究。
DOI：
10.1016/0167-4889(91)90038-y
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Degani,H
中科院分区：
文献类型：
--
作者：
Ronen,SM;Rushkin,E;Degani,H
用~(31)P和~(13)C-核磁共振仪测定了人乳腺癌细胞T47D Clone 11生长为小球体(150μm)时胆碱和乙醇胺掺入动力学。用1，2-13C标记的胆碱或1，2-13C标记的乙醇胺(0.028 mm)将球体注入光谱仪内，监测标记的磷酰胆碱(PC)或磷乙醇胺(PE)的积聚。或者，用13C预先标记PC和GPC池，并监测标记的减少。记录31P谱，从中可以确定总体能量状态以及总的池大小。ATP含量为8±1fmol/cell，PC池和PE池总大小分别为16和14fmol/cell。在没有添加乙醇胺的情况下，PC在24 h内增加了50%或保持不变，而在添加乙醇胺的情况下，PE在30 h内增加了2倍，这表明这与介质中前体的浓度有关。31P和13C数据得到相似的动力学结果：两种酶的速率均为1.0fmol/cell/h，CTP：磷酸胆碱胞苷转移酶和CTP：磷酸乙醇胺激酶的速率常数均为0.06h−1。胆碱掺入动力学在0.028 mM乙醇胺存在下没有改变，表明它们存在非竞争途径。
31P and13C-NMR were used to determine the kinetics of choline and ethanolamine incorporation in T47D clone 11 human breast cancer cells grown as small (150 μm) spheroids. Spheroids were perfused inside the spectrometer with 1,2-13C-labeled choline or 1,2-13C-labeled ethanolamine (0.028 mM) and the buildup of labeled phosphorylcholine (PC) or phosphorylethanolamine (PE) was monitored. Alternatively the PC and GPC pools were prelabeled with13C and the reduction of label was monitored.31P spectra were recorded from which the overall energetic status as well as total pool sizes could be determined. The ATP content was 8 ± 1 fmol/cell, and the total PC and PE pool sizes were 16 and 14 fmol/cell, respectively. PC either increased by 50% over 24 h or remained constant, while PE remained constant in medium without added ethanolamine but increased 2-fold within 30 h in medium containing ethanolamine, indicating a dependence on precursor concentration in the medium. The31P and13C data yielded similar kinetic results: the rate of the enzymes phosphocholine kinase and phosphoethanolanine kinase were both on the order of 1.0 fmol/cell per h, and the rate constants for CTP:phosphocholine cytidyltransferase and CTP:phosphoethanolamine kinase were 0.06 h−1for both enzymes. The kinetics of choline incorporation did not alter in the presence of 0.028 mM ethanolamine indicating that they have non-competing pathways.