Serum Metabolite Profiling of Human Colorectal Cancer Using GC−TOFMS and UPLC−QTOFMS

Abstract

Colorectal carcinogenesis involves the overexpression of many immediate-early response genes associated with growth and inflammation, which significantly alters downstream protein synthesis and small-molecule metabolite production. We have performed a serum metabolic analysis to test the hypothesis that the distinct metabolite profiles of malignant tumors are reflected in biofluids. In this study, we have analyzed the serum metabolites from 64 colorectal cancer (CRC) patients and 65 healthy controls using gas chromatography time-of-flight mass spectrometry (GC-TOFMS) and Acquity ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (Acquity UPLC-QTOFMS). Orthogonal partial least-squares discriminate analysis (OPLS-DA) models generated from GC-TOFMS and UPLC-QTOFMS metabolic profile data showed robust discrimination from CRC patients and healthy controls. A total of 33 differential metabolites were identified using these two analytical platforms, five of which were detected in both instruments. These metabolites potentially reveal perturbation of glycolysis, arginine and proline metabolism, fatty acid metabolism and oleamide metabolism, associated with CRC morbidity. These results suggest that serum metabolic profiling has great potential in detecting CRC and helping to understand its underlying mechanisms.

UPLC-based metabonomic applications for discovering biomarkers of diseases in clinical chemistry

“…GC-MS and UPLC-MS-based metabonomics developed and applied on the serum from CRC patients revealed perturbation of glycolysis, arginine and proline metabolism, fatty acid metabolism and oleamide metabolism and its association with CRC morbidity [65]. Tricarboxylic acid cycle, urea cycle, glutamine, fatty acids, and gut flora metabolism were disturbed [66], which are consistent with previous findings [67].…”

Tissue metabolomic fingerprinting reveals metabolic disorders associated with human gastric cancer morbidity

“…The use of GC/MS in the metabolic profiling of esophageal cancer (19), colorectal cancer (10,26,27,35), liver cancer (16), bladder cancer (30) and ovarian cancer (32) has been reported, while few studies have investigated the metabolic profiles of GC tissue. This study explored the metabolic fingerprinting of GC tumor specimens and their matched normal mucosa.…”

“…OpLs-DA modeling for the observation of overall separation trends between cancer and non-cancer cohorts has been adopted in human colorectal and bladder cancer (26,27,30,35). previous studies indicate that the supervised model is a robust model for discrimination.…”

“…Fifty milligrams of each tissue was transferred to 2.0-ml eppendorf tubes and lyophilized and extracted with 1.0 ml of a mixture of methanol/ chloroform (3:1, v/v) and vortexed for 30 sec (26). After storing for 10 min at -20˚C, the samples were subsequently centrifuged at 12,000 x g for 10 min at 4˚C, and 200 µl of the supernatant was collected separately from each sample and transferred to a 1.5-ml eppendorf tube.…”

NMR-based Metabolomic Analysis of Human Bladder Cancer

“…4 Metabolomics, as a combination of spectroscopic measurement of biological samples with multivariate data analysis, has been employed extensively to address the diagnosis and evaluation of cancer patients, and to identify potential biomarkers. [5][6][7][8] Another related term "metabonomics" was initially defined as "the quantitative measurement of the multi-parametric metabolic response of living systems to pathophysiological stimuli or genetic modifications". 9 The definition of metabolomics is evolving rapidly towards its "brother"-metabonomics.…”