1H NMR-Based Metabonomic Investigation Of The Liver Cancers' Urine

Globally,liver cancer is a malignant liver disease,ranking sixth among common malignancies and third among fatal cancers.Early diagnosis and treatment of liver cancer will obviously improve the survival of patients and improve the survival rate of patients with liver cancer,but there is no specific early diagnosis method.Metabonomics focuses on a systems-based study of all small molecule metabolites in the body.One of the major applications of metabonomic is disease diagnosis.Since the metabolome changes in the body with pathological,the analysis of these metabolites can help us understand the metabolic pathways of the metabolites and the disease process,find the biomarker(s)and could be supplementary means in clinical diagnosis.Nuclear Magnetic Resonance(NMR)was used to investigate the different metabolites between urine samples from patients with liver cancer and normal controls.We discriminated the liver cancer subjects from the healthy controls by their urinary metabolic profiles.Combined with supervised pattern recognition and univariate statistical method,it can effectively find out the differential metabolites contributed for the separation between the patients with liver cancer and normal controls derived from urine sample.A total of 12 identified potential biomarkers were found related to the disease progression.A panel of metabolite markers composed of 3 metabolites was able to discriminate the liver cancer subjects from their healthy counterparts.A receiver operating characteristic curve(ROC)analysis of these markers resulted in an area under the receiver operating characteristic curve(AUC)of 0.972.The results may provide an insight into the understanding of the biochemical network and pathway in liver cancer.This thesis consists of five parts as follows:The first part introduces the background and significance of this topic,the reported literatures on the metabolism of liver-related diseases were combed.We determines that only Shariff and others had studied the urine samples of liver cancer caused by hepatitis C virus,thus confirming the importance of finding the key metabolic markers of liver cancer urine samples as early diagnosis.The work of the second part is mainly to establish the amino acid database based on NMR analysis method.An amino acid standard solution was prepared with heavy water phosphate buffer solution,and hydrogen spectrums were collected to form a small database.The special property of the nuclear magnetic resonance was used to identify the amino acid components in the samples.This makes a good preparation for the identification of urine metabolites in later chapters.The work of the third part is mainly to establish the 1H-NMR analysis method of urine.Through continuous experiment and constant adjustment of parameter setting,the volume of 300 ?L urine sample is suitable.The specific parameters and processing parameters are specified in the non-rotating state.The work of the four part is mainly to investigate the different metabolic profiles of patients with liver cancer and normal controls using NMR method in the urinary metabonomic study.The results showed that urine metabolic profiles of patients with liver cancer and normal controls significantly differ from each other.12 most important markers were found,indicating great potential for early diagnosis of liver cancer.Our findings confirm a distinct urinary metabolic profile of patients with liver cancer characterized by altered levels of many metabolites involved in alpha-glucose,beta-glucose,taurine,valine,2-hydroxybutyric acid,formic acid,creatinine,creatine,and uric acid,butyric acid,citric acid and dimethylamine.A panel of metabolite markers composed of 3 metabolites(alpha-glucose,beta-glucose and creatinine)selected by univariate statistical method,was able to discriminate patients with liver cancer from normal controls,with an AUC of 0.972.The last part is to summarize the shortage of the experiment and the prospect of the future.