Serum and Urine Metabolite Profiling Reveals Potential Biomarkers of Human Hepatocellular Carcinoma

Tianlu Chen,Guoxiang Xie,Xiaoying Wang,Xin Qi,Xiaojiao Zheng,Xiaoyan Wang,Jia Fan,Yu Cao,Ping Liu,Lisa X Xu,Mingming Su,Yun Yen,Wei Jia,Yunping Qiu

doi:10.1074/mcp.m110.004945

Abstract

Hepatocellular carcinoma (HCC) is a common malignancy in the world with high morbidity and mortality rate. Identification of novel biomarkers in HCC remains impeded primarily because of the heterogeneity of the disease in clinical presentations as well as the pathophysiological variations derived from underlying conditions such as cirrhosis and steatohepatitis. The aim of this study is to search for potential metabolite biomarkers of human HCC using serum and urine metabolomics approach. Sera and urine samples were collected from patients with HCC (n = 82), benign liver tumor patients (n = 24), and healthy controls (n = 71). Metabolite profiling was performed by gas chromatography time-of-flight mass spectrometry and ultra performance liquid chromatography-quadrupole time of flight mass spectrometry in conjunction with univariate and multivariate statistical analyses. Forty three serum metabolites and 31 urinary metabolites were identified in HCC patients involving several key metabolic pathways such as bile acids, free fatty acids, glycolysis, urea cycle, and methionine metabolism. Differentially expressed metabolites in HCC subjects, such as bile acids, histidine, and inosine are of great statistical significance and high fold changes, which warrant further validation as potential biomarkers for HCC. However, alterations of several bile acids seem to be affected by the condition of liver cirrhosis and hepatitis. Quantitative measurement and comparison of seven bile acids among benign liver tumor patients with liver cirrhosis and hepatitis, HCC patients with liver cirrhosis and hepatitis, HCC patients without liver cirrhosis and hepatitis, and healthy controls revealed that the abnormal levels of glycochenodeoxycholic acid, glycocholic acid, taurocholic acid, and chenodeoxycholic acid are associated with liver cirrhosis and hepatitis. HCC patients with alpha fetoprotein values lower than 20 ng/ml was successfully differentiated from healthy controls with an accuracy of 100% using a panel of metabolite markers. Our work shows that metabolomic profiling approach is a promising screening tool for the diagnosis and stratification of HCC patients.

Highlights

From the ‡Ministry of Education Key Laboratory of Systems Biomedicine, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China; §Department of Nutrition, University of North Carolina at Greensboro, North Carolina Research Campus, Kannapolis, North Carolina 28081, USA; ¶Zhongshan Hospital, Shanghai 200030, China; ʈSchool of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; **David H
We have recently demonstrated that a combination of gas chromatography time-offlight mass spectrometry (GC-TOFMS) and ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOFMS) significantly increased the number of serum metabolite markers identified in a clinical metabolomic study of colorectal cancer [23]
A total of 324 peaks were obtained from GC-TOFMS spectra, whereas 2626 peaks were obtained from UPLC-QTOFMS ESϩ mode and 925 peaks obtained from ESmode

Summary

Introduction

From the ‡Ministry of Education Key Laboratory of Systems Biomedicine, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China; §Department of Nutrition, University of North Carolina at Greensboro, North Carolina Research Campus, Kannapolis, North Carolina 28081, USA; ¶Zhongshan Hospital, Shanghai 200030, China; ʈSchool of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China; **David H. And accurate diagnosis of HCC is of central importance for timely treatment and five-year survival rate Genomics and proteomics have merged as biochemical profiling tools to provide important insight into the biology of various cancers [11] These profiling approaches focus on upstream genetic and protein variations, metabolomics captures the global metabolic changes that occur in response to pathological, environmental or lifestyle factors [12]. We have recently demonstrated that a combination of gas chromatography time-offlight mass spectrometry (GC-TOFMS) and ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOFMS) significantly increased the number of serum metabolite markers identified in a clinical metabolomic study of colorectal cancer [23]

Objectives

Results

Conclusion