Non-targeted And Targeted Human Serum Metabolomics Of Hepatocellular Carcinoma

The liver,regarded as metabolic center of human,involves in the progress of substance metabolism.Hepatocellular carcinoma(HCC)caused by internal or external factors could lead to changes of metabolic fluxes and metabolites concentration in human serum.Metabolomics is an analytical technique with high sensitivity,high resolution and high throughput,which can sensitively detect the changes of metabolite concentrations in HCC serum.At the present,the metabolite numbers in human serum detected by metabolomics of hepatocellular carcinoma are few due to the single methods.In this study,a more sensitive metabolomics method was optimized and established to get more metabolites,screen potential biomarkers in HCC serum and contribute to understanding the pathological mechanism.First of all,in order to explore HCC serum metabolites,our study had developed and validated a non-targeted metabolomics method on ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS)instrument with high resolution.After liquid-liquid extraction(LLE),the chromatographic separation had been achieved by T3 and C18 columns to separate the hydrophilic and lipophilic samples,independently.The results showed that 16 different metabolites were associated with HCC.Among them,the concentrations of palmitoyl ethanolamide(PEA)in HCC serum increased significantly compared to the healthy group.Based on the above results,a targeted metabolomics analysis method was then established and validated to detect 25 endocannabinoids(ECs)by using ultra performance liquid chromatography tandem triple-quadrupole mass spectrometry(UPLC-QQQ/MS)technique with high sensitivity.The results showed that 11 kinds of ECs in HCC serum,including arachidonoyl ethanolamide(AEA),PEA,oleoyl ethanolamide(OEA),linoleoyl ethanolamide(LEA),docosahexaenoyl ethanolamide(DHEA),etc,changed obviously compared to those in the healthy group.In summary,a comprehensive method using non-targeted and targeted metabolomics analysis technique to discover potential biomarkers had been developed.It revealed that ECs and corresponding metabolizing enzymes played an important role in medicine targeted therapy of HCC,providing a new way to study HCC mechanism at the molecular level and technical supports for basic research,translational medicine and pharmacy.