The Serum Metabolic Profiling On Different Stages Of BCLC Of HCC Patients Related With Hepatitis B

Objective To explore the characteristic ions in serum about different stages of BCLC of HCC patients related with hepatitis B by the UPLC- MS platform, and to establish models that could distinguish the stages. And then analyze these substances involved in the metabolic pathways, aimed at evaluating their values for diagnosing HCC with hepatitis B and monitoring the progress. So it could provide help for clinical judgment and finding new treat targets for individualized diagnosis and treatment.Methods The serum were sampled from 75 HCC patients related hepatitis B virus, who were diagnosed originally in the liver medicine and surgery departments of Tianjin Third Central Hospital from November, 2013 to January, 2015. They were divided into three groups according to the Barcelona liver cancer staging criteria:(1) early group 26 cases(group A),who can be treated radically;(2) middle group 23 cases(group B), who can be treated by arterial chemo embolization;(3) late group 26 cases(group C), who were only accepted symptomatic treatment. The control group included 20 health volunteers and 20 patients with HBV-induced cirrhosis. All the samples were tested by the Ultra-performance liquid chromatography combined with mass spectrometry(UPLC-MS), data was pretreated and normalized by the software of Mzmine 2.0. Orthogonal partial least squares discriminant analysis(OPLS-DA) model were constructed from the tissue metabolite profile data, and variable importance in projection(VIP) and VIP confidence intervals were then used to select potential biomarkers. Then, independent sample-nonparametric test was used to make the final screening, excluding the variables with P>0.05 from these selected metabolite biomarkers. And then, the irons would be identified. At last, ROC curve was used to evaluate the effectiveness of these irons which could diagnose the different BCLC stages of HCC.Results(1)20 control samples were tested, a PCA score discriminant model diagram containing a main component was established(R2X =16.4%, Q2 =5.99%). All the control samples were in the?±2SD, there were 18 samples meet the requirements of quality control materials with a pass rate of 90%.(2)Serum metabonomic PCA and OPLS-DA models were established successfully to diagnose different BCLC stages of HCC related HBV, R2X=67.2%, R2Y=82%, Q2Y=61.1%.(3)51 metabolites which had statistical differences between A group and C group were selected from OPLS-DA models, there were 20 metabolites have statistical differences among groups: 6 in cirrhosis group and HCC group, 7 in healthy group and HCC group,2 inevery stage of HCC, 9 in A group and B、C groups and 8 in C group and A、B groups.(4)Above-mentioned 20 kinds of characteristics of metabolic ions highly associated with liver tumor development were identified, mainly include lysophosphatidylcholine、amino acids、glycine acid and vaccenic acid.(5) The area under the ROC curve of Lyso PC(18:2(9Z,12Z))、Lyso PC(P-16:0)、Asparaginyl- Proli- ne and vaccenic acid between the compare of HCC and cirrhosis were all bigger than AFP, having better diagnosis ability to HCC especially to early HCC. And the area of L-Aspartyl-4-phosphate and Lyso PC(20:5(5Z,8Z,11 Z,14Z,17Z)) in A、B groups compare were bigger than AFP, but were smaller in the compare of B、C groups.Conclusions This study successed in screening metabolic ions which can diagnose HCC and reflect progress of tumor and at the same time they had high diagnostic efficiency based on the UPLC-MS platform and multivariate statistical analysis. To understand the trends of these endogenous metabolites in the patients of HCC related with HBV would play a good role in diagnosing diseases and monitoring the development course of diseases. And it could provide strong support for the development of clinical "individualized" treatment program.