8-Hydroxyguanosine as a possible RNA oxidative modification marker in urine from colorectal cancer patients: Evaluation by ultra performance liquid chromatography-tandem mass spectrometry
8-Hydroxyguanosine as a possible RNA oxidative modification marker in urine from colorectal cancer patients: Evaluation by ultra performance liquid chromatography-tandem mass spectrometry
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DOI：
10.1016/j.jchromb.2019.121931
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发表时间：
2020-01-01
影响因子：
3
通讯作者：
Zheng, Shu
中科院分区：
文献类型：
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作者：
Guo, Cheng;Chen, Qin;Zheng, Shu
RNA氧化损伤已被发现与多种疾病有关，8-羟基鸟苷（8-OHG）是RNA氧化修饰的典型标志物。这种鸟苷修饰是一种新兴的疾病检测生物标志物，人类尿液中8-羟色胺的测定受到青睐，因为它对患者无创。然而，由于其在质谱中的电离效率较差，且在尿液中痕量，因此对其进行精确定量仍然具有挑战性。本研究建立了一种快速、准确、灵敏、可靠的固相萃取（SPE）联合同位素稀释超高效液相色谱-串联质谱（UPLC-MS/MS）检测人尿中氧化性RNA修饰的方法。检出限可达1.5 fmol，日间（1.8-3.3%）和日间（0.6-1.2%）分析精密度良好。采用HLB药筒对尿液进行预处理，在三个加标水平下回收率为87.5-107.2%，令人满意。采用该方法对65例结直肠癌（CRC）患者和76名健康志愿者尿液中的8-羟色胺进行了定量分析。结直肠癌患者和健康对照组尿8-OHG测定值分别为1.91 +/- 0.63 nmol/mmol肌酐和1.33 +/- 0.35 nmol/mmol肌酐。我们发现CRC患者尿液中8-OHG含量升高，这意味着这种氧化RNA修饰标记物可以作为CRC早期筛查的潜在无创指标。此外，本研究将有助于研究氧化应激对结直肠癌形成和发展的影响。
Oxidative RNA damage has been found to be associated with a variety of diseases, and 8-hydroxyguanosine (8-OHG) is a typical marker of oxidative modification of RNA. This guanosine modification is an emerging biomarker for disease detection and determination of 8-OHG in human urine is favored because it is noninvasive to patients. However, due to its poor ionization efficiency in mass spectrometry and trace amount in urine, accurate quantification of this modified nucleoside is still challenging. Herein, a rapid, accurate, sensitive and robust method using solid-phase extraction (SPE) combined with isotope dilution ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed for detection of this oxidative RNA modification in human urine. The limit of detection can reach 1.5 fmol and the method exhibits good precision on intra-day (1.8-3.3%) and inter-day (0.6-1.2%) analyses. Satisfactory recovery (87.5-107.2%) at three spiked levels was achieved by using HLB cartridge for urine pretreatment. Using this method, we quantified 8-OHG in urine from 65 colorectal cancer (CRC) patients and 76 healthy volunteers. The measured level of urinary 8-OHG for CRC patients and healthy controls is 1.91 +/- 0.63 nmol/mmol creatinine and 1.33 +/- 0.35 nmol/mmol creatinine, respectively. We found the content of 8-OHG in urine was raised in CRC patients patients, implying this oxidative RNA modification marker could act as a potential noninvasive indicator for early screening of CRC. In addition, this study will make contributions to the investigations of the influences of oxidative stress on the formation and development of CRC.