A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels.
A LC-MS/MS method for the analysis of intracellular nucleoside triphosphate levels.
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DOI：
10.1007/s11095-009-9863-9
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发表时间：
2009-06
影响因子：
3.7
通讯作者：
Chan, Kenneth K.
中科院分区：
文献类型：
--
作者：
Chen, Ping;Liu, Zhongfa;Liu, Shujun;Xie, Zhiliang;Aimiuwu, Josephine;Pang, Jiuxia;Klisovic, Rebecca;Blum, William;Grever, Michael R.;Marcucci, Guido;Chan, Kenneth K.
同时定量细胞内三磷酸核苷 (NTP) 和三磷酸脱氧核苷 (dNTP) 库，并评估干扰白血病细胞中核糖核苷酸还原酶 (RNR) 表达而产生的变化。 HPLC-MS/MS 系统用于量化细胞内 NTP 和 dNTP 库。该测定在定量下限 (LLOQ) 50 nM 和 10 μMin 细胞裂解液之间呈线性。 dNTP 的日内变异系数（CV，n=5）在 LLOQ 处为 12.0–18.0%，在 500 至 5,000 nM 之间为 3.0–9.0%，NTP 为 8.0–15.0% 和 2.0–6.0%。 dNTP 的日间 CV (n=5) 为 9.0–13.0% 和 3.0–11.0%，NTP 的日间 CV 为 9.0–13.0% 和 3.0–6.0%。 NTP 和 dNTP 的日内准确度值为 93.0–119.0%。 ATP 与 dGTP 重叠，并将它们作为复合物进行分析。该方法用于测量暴露于 RNR 反义 GTI-2040 的五种白血病细胞系中的基础细胞内 dNTP/NTP。药物治疗后，MV4-11 和 K562 细胞中 dCTP 和 dATP 水平显着降低。此外，还检测到接受 GTI-2040 治疗的患者骨髓样本中 dNTP/NTP 水平的扰动。该方法提供了测量细胞和临床样本中细胞内 dNTP/NTP 水平的实用工具。
To simultaneously quantify intracellular nucleoside triphosphate (NTP) and deoxynucleoside triphosphate (dNTP) pools and to assess their changes produced by interfering with ribonucleotide reductase (RNR) expression in leukemia cells. A HPLC-MS/MS system was used to quantify intracellular NTP and dNTP pools. The assay was linear between 50 nM, the lower limit of quantification (LLOQ), and 10 μMin cell lysate. The within-day coefficients of variation (CVs, n=5) were found to be 12.0–18.0% at the LLOQ and 3.0–9.0% between 500 and 5,000 nM for dNTPs and 8.0–15.0% and 2.0–6.0% for NTPs. The between-day CVs (n=5) were 9.0–13.0% and 3.0–11.0% for dNTPs and 9.0–13.0% and 3.0–6.0% for NTPs. The within-day accuracy values were 93.0–119.0% for both NTPs and dNTPs. ATP overlapped with dGTP and they were analyzed as a composite. This method was applied to measure basal intracellular dNTPs/NTPs in five leukemia cell lines exposed to the RNR antisense GTI-2040. Following drug treatment, dCTP and dATP levels were found to decrease significantly in MV4-11 and K562 cells. Additionally, perturbation of dNTP/NTP levels in bone marrow sample of a patient treated with GTI-2040 was detected. This method provides a practical tool to measure intracellular dNTP/NTP levels in cells and clinical samples.
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影响因子：
64.5
作者：
Chandra, Dhyan;Bratton, Shawn B.;Tang, Dean G.
通讯作者：
Tang, Dean G.
影响因子：
3.7
作者：
FERRARO, P;BIANCHI, V;CELOTTI, L
通讯作者：
CELOTTI, L
高效液相色谱法分析正常细胞系和肿瘤细胞系的细胞内三磷酸核苷水平
DOI：
10.1016/s1570-0232(02)00780-8
发表时间：
2003-01-25
影响因子：
3
作者：
Huang, D;Zhang, YH;Chen, XG
通讯作者：
Chen, XG
DOI：
10.1016/s0378-4347(00)00619-8
发表时间：
2001-04-25
期刊：
JOURNAL OF CHROMATOGRAPHY B
影响因子：
--
作者：
Fung, EN;Cai, ZW;Sinhababu, AK
通讯作者：
Sinhababu, AK
影响因子：
8.5
作者：
CROSS, DR;MILLER, BJ;JAMES, SJ
通讯作者：
JAMES, SJ