Inhibition of tumor cell ribonucleotide reductase by macrophage-derived nitric oxide.
Inhibition of tumor cell ribonucleotide reductase by macrophage-derived nitric oxide.
复制标题
DOI：
10.1084/jem.174.4.761
复制
发表时间：
1991-10-01
期刊：
影响因子：
--
通讯作者：
Nathan CF
中科院分区：
文献类型：
--
作者：
Kwon NS;Stuehr DJ;Nathan CF
巨噬细胞来源的一氧化氮（NO）对肿瘤细胞和微生物病原体具有细胞抑制作用。我们测试了一氧化氮细胞抑制作用的一个分子靶标是否可能是核糖核苷酸还原酶（RR），一种DNA合成中的限速酶。以浓度依赖的方式，活化巨噬细胞的NO气体和裂解物产生相当数量的NO，导致L1210小鼠淋巴瘤细胞部分纯化的RR受到相同程度的抑制。非活化巨噬细胞的裂解物不产生NO，无抑制作用。对于活化的巨噬细胞的裂解物，通过省略l-精氨酸或添加NO合成酶抑制剂二苯乙烯酮、N - omega-甲基- l-精氨酸或N - omega-氨基- l-精氨酸来保护RR。l -精氨酸，而不是D-精氨酸，可以消除N - ω -氨基- l -精氨酸的保护作用。抗胰胆管炎的原型药理学抑制剂是羟基脲。它的结构与N - omega-羟基- l -精氨酸（NO合成酶的反应中间体）相似，这促使我们测试羟基脲是否能生成NO。在H2O2和CuSO4存在下，羟基脲生成NO的好氧反应产物NO2-/NO3-。通过气相色谱/质谱分析发现，加入morpholine阻断了羟基脲生成NO2-/NO3，并导致了nitrosomorpholine的生成。因此，羟基脲可以产生类似no的亚硝化物。L1210细胞的DNA合成被活化的巨噬细胞或羟基脲完全抑制，在两种情况下，通过提供脱氧核糖核苷来绕过RR中的阻滞，可以部分恢复到相同程度。因此，NO气体和活化的巨噬细胞裂解物产生的NO均抑制肿瘤细胞的RR。RR抑制剂羟基脲也能产生no样物质。同样，在活化的巨噬细胞或羟基脲存在的情况下，脱氧核糖核苷对肿瘤细胞DNA合成的部分恢复表明，活化的巨噬细胞和羟基脲的细胞抑制具有相似的机制，包括但可能不限于抑制RR。
Macrophage-derived nitric oxide (NO) is cytostatic to tumor cells and microbial pathogens. We tested whether one molecular target for the cytostatic action of NO may be ribonucleotide reductase (RR), a rate- limiting enzyme in DNA synthesis. In a concentration-dependent manner, NO gas and lysates of activated macrophages that generated comparable amounts of NO led to the same degree of inhibition of partially purified RR from L1210 mouse lymphoma cells. Lysates from nonactivated macrophages, which do not produce NO, were noninhibitory. With lysates from activated macrophages, RR was protected by omitting L-arginine or by adding the NO synthase inhibitors diphenyleneiodonium, N omega- methyl-L-arginine, or N omega-amino-L-arginine. L-Arginine, but not D- arginine, abolished the protective effect of N omega-amino-L-arginine. The prototypic pharmacologic inhibitor of RR is hydroxyurea. Its structural resemblance to N omega-hydroxy-L-arginine, a reaction intermediate of NO synthase, prompted us to test if hydroxyurea can generate NO. In the presence of H2O2 and CuSO4, hydroxyurea produced NO2-/NO3-, aerobic reaction products of NO. Addition of morpholine blocked NO2-/NO3- generation from hydroxyurea and led to formation of nitrosomorpholine, as detected by gas chromatography/mass spectrometry. Thus, hydroxyurea can produce an NO-like, nitrosating rectant. L1210 cell DNA synthesis was inhibited completely by activated macrophages or by hydroxyurea, and was partially restored to the same degree in both settings by providing deoxyribonucleosides to bypass the block in RR. Thus, both NO gas and NO generated by activated macrophage lysates inhibit tumor cell RR. The RR inhibitor hydroxyurea can also generate an NO-like species. Similar, partial restoration of tumor cell DNA synthesis by deoxyribonucleosides in the presence of activated macrophages or hydroxyurea suggests that cytostasis by activated macrophages and by hydroxyurea has comparable mechanisms, including, but probably not limited to, inhibition of RR.