NMR and LC/MS-based global metabolomics to identify serum biomarkers differentiating hepatocellular carcinoma from liver cirrhosis

Abstract

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. However, current biomarkers that discriminate HCC from liver cirrhosis (LC) are important but are limited. More reliable biomarkers for HCC diagnosis are therefore needed. Serum from HCC patients, LC patients and healthy volunteers were analyzed using NMR and LC/MS-based approach in conjunction with random forest (RF) analysis to discriminate their serum metabolic profiles. Thirty-two potential biomarkers have been identified, and the feasibility of using these biomarkers for the diagnosis of HCC was evaluated, where 100% sensitivity was achieved in detecting HCC patients even with AFP values lower than 20 ng/mL. The metabolic alterations induced by HCC showed perturbations in synthesis of ketone bodies, citrate cycle, phospholipid metabolism, sphingolipid metabolism, fatty acid oxidation, amino acid catabolism and bile acid metabolism in HCC patients. Our results suggested that these potential biomarkers identified appeared to have diagnostic and/or prognostic values for HCC, which deserve to be further investigated. In addition, it also suggested that RF is a classification algorithm well suited for selection of biologically relevant features in metabolomics.Hepatocellular carcinoma (HCC) is the most common intraabdominal malignancies and the third leading cause of cancer-related death worldwide, with a <7% 5-year survival rate.

Identification of serum biomarkers associated with hepatitis B virus-related hepatocellular carcinoma and liver cirrhosis using mass-spectrometry-based metabolomics

“…It is known that LysoPCs and LysoPEs regulate a variety of biological processes including cell proliferation, tumor cell invasiveness, and inflammation (Ressom et al 2012). The Identification of serum biomarkers associated with hepatitis B virus-related hepatocellular… down-regulation of them may mainly result from rapid membrane PC and PE turnover during the liver injury or malignant regeneration (Liu et al 2014). Acylcarnitines (carnitine-fatty acid esters) are essential intermediates in lipid metabolism, which facilitate transportation of longchain fatty acids into mitochondria for beta-oxidation to generate adenosine triphosphate (ATP), assist in the removal of short-and medium-chain fatty acids out of mitochondria, and maintain membrane phospholipid turnover (Sharma and Black 2009).…”

Lipid Metabolism in Liver Cancer

“…This theme will be further explored in the next section. As same Lipid Metabolism in Liver Cancer http://dx.doi.org/10.5772/64993as the intermediate liver diseases, HCC maintained the aforementioned "core metabolic phenotype" of bile acid and phospholipid perturbation [66,71,[73][74][75]. Some findings also demonstrated similar liver impairments of ammonium detoxication and amino acid metabolism in HCC as in cirrhosis [66,72,73,76].…”

“…This phonotype was evidenced by the following metabolic changes including the decrease in glucose, citrate, and glycerol 3-phosphate but the increase in lactic acid and pyruvate [64][65][66][67][68][69]. On the other hand, the common induction of beta-oxidation was exemplified by the elevation of 2-oxoglutarate and reduction of free fatty acids, carnitine and carnitine esters [61,64,66,68,[70][71][72][73]. This theme will be further explored in the next section.…”

“…One Chinese study using a panel of metabolic markers (formate, phytosphingosine, and 3α,6α,7α,12α-tetrahydroxy-5β-cholan-24-oleic acid) achieved high accuracy (area under curve (AUC): 0.995-1.000, sensitivity: 100%, and specificity: 94.7-100%) in diagnosing HCC patients with low alpha-fetoprotein (a commonly accepted but mediocre biomarker to differentiate HCC from non-tumor liver at a cut-off value of 20 ng/ml) [73]. Another Chinese study, which used tryptophan, glutamine, and 2-hydroxybutyric acid as biomarkers in 183 human serum, also achieved high accuracy (AUC: 0.969-0.990) in both training and validation dataset [69].…”

Identification of Cyclase-Associated Protein-2 as a Novel Biomarker for Early- Stage Hepatocellular Carcinoma

“…Thus, the early detection of HCC is an important and crucial goal for all researchers. The early diagnosis and prognostic prediction of HCC are much difficult due to the coexistence of liver cirrhosis and inflammation in HCC [9]. Unfortunately, alpha fetoprotein (AFP) which was the golden marker of HCC lacks both sensitivity and specificity, has limited performance especially in early-stage HCCs and is no longer recommended by international surveillance guidelines [10].…”