Inhibition of glutamine synthetase decreases proliferation of cultured rat intestinal epithelial cells

被引:46
|
作者
DeMarco, V [1 ]
Dyess, K
Strauss, D
West, CM
Neu, J
机构
[1] Univ Florida, Coll Med, Dept Pediat, Div Neonatol, Gainesville, FL 32610 USA
[2] Univ Florida, Coll Med, Dept Anat & Cell Biol, Gainesville, FL 32610 USA
来源
JOURNAL OF NUTRITION | 1999年 / 129卷 / 01期
关键词
endogenous glutamine production; IEC-6; cells; rats; small intestine; crypt cells;
D O I
10.1093/jn/129.1.57
中图分类号
R15 [营养卫生、食品卫生]; TS201 [基础科学];
学科分类号
100403 ;
摘要
The importance of glutamine synthetase (GS) for cell proliferation was examined in rat intestinal crypt cells (IEC-6) by inhibiting its activity with 10 mmol/L methionine sulfoximine (MS) at varying extracellular glutamine (Q) concentrations. In uninhibited cultures, cell number, protein, and DNA accumulation and synthesis showed a dependence on extracellular Q over a concentration range of 0.06 to 1.06 mmol/L, with apparent half-maximal responses of 0.46 mmol/L extracellular Q. In contrast, proliferation of GS-inhibited cultures required greater than or equal to 1.06 mmol/L extracellular Q, with an apparent half-maximal response of 2 mmol/L. MS inhibited GS activity >97% in extracts of washed cells and appeared to be specific because its effects on proliferation were overcome by 4.06 mmol/L Q and were reversible. The increased dependence of IEC-6 cells on extracellular Q when GS was inhibited suggests that Q derived from GS (GS-Q) contributes importantly to cell proliferation at physiologic levels of extracellular Q (0.6 mmol/L). The unexpectedly high concentration of extracellular Q required to rescue maximal proliferation during OS-inhibition, relative to a reported K-m for Q-transport into the cell, indicates that intracellular Q derived from the extracellular medium (exo-Q) is inefficiently utilized. In a previous study, we found that GS-protein and mRNA are concentrated in the proliferative crypt region of the small intestine in vivo, and predicted that GS activity is important for crypt cell proliferation. Here, we show that enzyme activity is important for cell proliferation at physiologic concentrations of Q in this cell culture model. Finally, we speculate that exo-Q and GS-Q are utilized differently in the cell.
引用
收藏
页码:57 / 62
页数:6
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