Development of Urinary Pseudotargeted LC-MS-Based Metabolomics Method and Its Application in Hepatocellular Carcinoma Biomarker Discovery
Development of Urinary Pseudotargeted LC-MS-Based Metabolomics Method and Its Application in Hepatocellular Carcinoma Biomarker Discovery
复制标题
基于尿液伪靶向LC-MS的代谢组学方法的发展及其在肝细胞癌生物标志物发现中的应用
DOI：
10.1021/pr500973d
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发表时间：
2015-02-01
影响因子：
4.4
通讯作者：
Yao, Zhenzhen
中科院分区：
文献类型：
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作者：
Shao, Yaping;Zhu, Bin;Yao, Zhenzhen
肝细胞癌（Hepatocellular carcinoma，HCC）是导致癌症相关死亡的主要恶性肿瘤之一。发现有效的肝癌诊断标志物是一个迫切的需求。为了鉴定潜在的代谢物生物标志物，我们开发了一种基于液相色谱-混合三重四极杆线性离子阱质谱（LC-QTRAP MS）的尿液假靶向方法。与非靶向方法相比，假靶向方法可以获得更好的数据质量，有利于差异代谢物的发现。所建立的方法被应用于肝硬化（CIR）和肝癌的调查。结果发现，肝病组尿核苷、胆汁酸、柠檬酸及多种氨基酸与对照组相比有明显变化，表现为肝脏嘌呤代谢、能量代谢和氨基酸代谢的异常。此外，一些代谢物，如环磷酸腺苷，谷氨酰胺，短链和中链酰基肉毒碱是HCC和CIR的差异代谢物。在二元Logistic回归的基础上，丁酰肉碱（肉毒碱C4：0）和海因-5-丙酸被定义为区分HCC和CIR的组合标志物。发现阶段和验证阶段样品的曲线下面积分别为0.786和0.773。这些数据表明，所建立的假靶向方法是代谢组学研究中靶向和非靶向方法的补充。
Hepatocellular carcinoma (HCC) is one of the pestilent malignancies leading to cancer-related death. Discovering effective biomarkers for HCC diagnosis is an urgent demand. To identify potential metabolite biomarkers, we developed a urinary pseudotargeted method based on liquid chromatography-hybrid triple quadrupole linear ion trap mass spectrometry (LC-QTRAP MS). Compared with nontargeted method, the pseudotargeted method can achieve better data quality, which benefits differential metabolites discovery. The established method was applied to cirrhosis (CIR) and HCC investigation. It was found that urinary nucleosides, bile acids, citric acid, and several amino acids were significantly changed in liver disease groups compared with the controls, featuring the dysregulation of purine metabolism, energy metabolism, and amino metabolism in liver diseases. Furthermore, some metabolites such as cyclic adenosine monophosphate, glutamine, and short- and medium-chain acylcarnitines were the differential metabolites of HCC and CIR. On the basis of binary logistic regression, butyrylcarnitine (carnitine C4:0) and hydantoin-5-propionic acid were defined as combinational markers to distinguish HCC from CIR. The area under curve was 0.786 and 0.773 for discovery stage and validation stage samples, respectively. These data show that the established pseudotargeted method is a complementary one of targeted and nontargeted methods for metabolomics study.