Stable Isotope Resolved Metabolomics Analysis of Ribonucleotide and RNA Metabolism in Human Lung Cancer Cells.
Stable Isotope Resolved Metabolomics Analysis of Ribonucleotide and RNA Metabolism in Human Lung Cancer Cells.
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DOI：
10.1007/s11306-011-0337-9
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发表时间：
2012-06
期刊：
影响因子：
3.6
通讯作者：
Lane, Andrew N.
中科院分区：
文献类型：
--
作者：
Fan, Teresa W-M.;Tan, Jinlian;McKinney, Martin M.;Lane, Andrew N.
我们开发了一种简单的基于核磁共振的方法，通过稳定同位素分解代谢组学（SIRM）来测定A549肺癌细胞中核苷酸的转换和RNA的结合。这种方法不需要对核苷酸进行化学降解或层析。在细胞生长过程中，游离的核糖核苷酸库迅速被新合成的核苷酸所取代。使用[U-13C]-葡萄糖和[U-13C，15N]-谷氨酰胺作为示踪剂，我们发现核苷酸核糖中的几乎所有碳都来自葡萄糖，而谷氨酰胺则优先于葡萄糖用于嘧啶环的生物合成，通过克雷布斯循环合成Asp。通过质子检测的15N核磁共振也证实了谷氨酰胺氨氮在嘌呤环的N3和N9位上的掺入。葡萄糖中的13C与总RNA的结合是细胞增殖过程中核苷酸的主要沉淀。本方法用于测定抗癌硒剂（甲基硒酸或MSA）对A549细胞的代谢作用。我们发现MSA抑制了核苷酸的转换和RNA的结合，这表明核苷酸代谢在MSA对癌细胞的毒性作用中起着重要作用。
We have developed a simple NMR-based method to determine the turnover of nucleotides and incorporation into RNA by stable isotope resolved metabolomics (SIRM) in A549 lung cancer cells. This method requires no chemical degradation of the nucleotides or chromatography. During cell growth, the free ribonucleotide pool is rapidly replaced by de novo synthesized nucleotides. Using [U-13C]-glucose and [U-13C,15N]-glutamine as tracers, we showed that virtually all of the carbons in the nucleotide riboses were derived from glucose, whereas glutamine was preferentially utilized over glucose for pyrimidine ring biosynthesis, via the synthesis of Asp through the Krebs cycle. Incorporation of the glutamine amido nitrogen into the N3 and N9 positions of the purine rings was also demonstrated by proton-detected 15N NMR. The incorporation of 13C from glucose into total RNA was measured and shown to be a major sink for the nucleotides during cell proliferation. This method was applied to determine the metabolic action of an anti-cancer selenium agent (methylseleninic acid or MSA) on A549 cells. We found that MSA inhibited nucleotide turnover and incorporation into RNA, implicating an important role of nucleotide metabolism in the toxic action of MSA on cancer cells.
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影响因子：
4.8
作者：
Lu, Xin;Bennet, Bryson;Kang, Yibin
通讯作者：
Kang, Yibin
影响因子：
3.6
作者：
Fan, Teresa W-M;Yuan, Peixiong;Lane, Andrew N.;Higashi, Richard M.;Wang, Yun;Hamidi, Anahita B.;Zhou, Rulun;Guitart, Xavier;Chen, Guang;Manji, Husseini K.;Kaddurah-Daouk, Rima
通讯作者：
Kaddurah-Daouk, Rima
影响因子：
3.3
作者：
Mendes, Ana C.;Caldeira, M. Madalena;Jones, John G.
通讯作者：
Jones, John G.
影响因子：
37.3
作者：
Fan TW;Lane AN;Higashi RM;Farag MA;Gao H;Bousamra M;Miller DM
通讯作者：
Miller DM
影响因子：
3.6
作者：
Beger, Richard D.;Hansen, Deborah K.;Schnackenberg, Laura K.;Cross, Brandie M.;Fatollahi, Javad J.;Lagunero, F. Tracy;Sarnyai, Zoltan;Boros, Laszlo G.
通讯作者：
Boros, Laszlo G.