Metabolomic Analysis of Liver Tissue from the VX2 Rabbit Model of Secondary Liver Tumors

Purpose. The incidence of liver neoplasms is rising in USA. The purpose of this study was to determine metabolic profiles of liver tissue during early cancer development. Methods. We used the rabbit VX2 model of liver tumors (LT) and a control group consisting of sham animals implanted with Gelfoam into their livers (LG). After two weeks from implantation, liver tissue from lobes with and without tumor was obtained from experimental animals (LT+/LT−) as well as liver tissue from controls (LG+/LG−). Peaks obtained by Gas Chromatography-Mass Spectrometry were subjected to identification. 56 metabolites were identified and their profiles compared between groups using principal component analysis (PCA) and a mixed-effect two-way ANOVA model. Results. Animals recovered from surgery uneventfully. Analyses identified a metabolite profile that significantly differs in experimental conditions after controlling the False Discovery Rate (FDR). 16 metabolites concentrations differed significantly when comparing samples from (LT+/LT−) to samples from (LG+/LG−) livers. A significant difference was also shown in 20 metabolites when comparing samples from (LT+) liver lobes to samples from (LT−) liver lobes. Conclusion. Normal liver tissue harboring malignancy had a distinct metabolic signature. The role of metabolic profiles on liver biopsies for the detection of early liver cancer remains to be determined.

[1]  S. Jonjić,et al.  Site-restricted persistent cytomegalovirus infection after selective long-term depletion of CD4+ T lymphocytes , 1989, The Journal of experimental medicine.

[2]  J. Garcia-conde,et al.  Serum metabolome analysis by 1H-NMR reveals differences between chronic lymphocytic leukaemia molecular subgroups , 2010, Leukemia.

[3]  R. Wahl,et al.  Targeting of VX2 Rabbit Liver Tumor by Selective Delivery of 3-Bromopyruvate: A Biodistribution and Survival Study , 2008, Journal of Pharmacology and Experimental Therapeutics.

[4]  D. Rosenberg,et al.  Roles of cPLA2α and arachidonic acid in cancer , 2006 .

[5]  Steven Mennerick,et al.  An integrated functional genomics and metabolomics approach for defining poor prognosis in human neuroendocrine cancers. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[6]  O. Warburg.  [Origin of cancer cells]. , 1956, Oncologia.

[7]  J. Sanabria,et al.  The Dynamics of Glutathione Species and Ophthalmate Concentrations in Plasma from the VX2 Rabbit Model of Secondary Liver Tumors , 2011, HPB surgery : a world journal of hepatic, pancreatic and biliary surgery.

[8]  Jean-Eudes J. Dazard,et al.  Joint adaptive mean-variance regularization and variance stabilization of high dimensional data , 2012, Comput. Stat. Data Anal..

[9]  Yong Yue,et al.  Metabonomic studies of human hepatocellular carcinoma using high-resolution magic-angle spinning 1H NMR spectroscopy in conjunction with multivariate data analysis. , 2007, Journal of proteome research.

[10]  Theodoros N. Arvanitis,et al.  Metabolic changes in flatfish hepatic tumours revealed by NMR-based metabolomics and metabolic correlation networks. , 2008, Journal of proteome research.

[11]  Z. Hall.  Cancer , 1906, The Hospital.

[12]  John D. Storey,et al.  Empirical Bayes Analysis of a Microarray Experiment , 2001 .

[13]  H. Dienes,et al.  Decreased hepatocyte membrane potential differences and GABAa‐β3 expression in human hepatocellular carcinoma , 2007, Hepatology.

[14]  Tao-Tao Liu,et al.  Metabolomic profiling of human urine in hepatocellular carcinoma patients using gas chromatography/mass spectrometry. , 2009, Analytica chimica acta.

[15]  C M Kendziorski,et al.  On parametric empirical Bayes methods for comparing multiple groups using replicated gene expression profiles , 2003, Statistics in medicine.

[16]  Scott F Lempka,et al.  Combined radiofrequency ablation and doxorubicin-eluting polymer implants for liver cancer treatment. , 2007, Journal of biomedical materials research. Part A.

[17]  Stefano Tiziani,et al.  Early stage diagnosis of oral cancer using 1H NMR-based metabolomics. , 2009, Neoplasia.

[18]  C. Dolea,et al.  World Health Organization , 1949, International Organization.

[19]  John R Griffiths,et al.  Metabolic changes detected by in vivo magnetic resonance studies of HEPA-1 wild-type tumors and tumors deficient in hypoxia-inducible factor-1beta (HIF-1beta): evidence of an anabolic role for the HIF-1 pathway. , 2002, Cancer research.

[20]  R. Kauppinen,et al.  Tumour metabolomics in animal models of human cancer. , 2007, Journal of proteome research.

[21]  P. Fortina,et al.  The reverse Warburg effect: Aerobic glycolysis in cancer associated fibroblasts and the tumor stroma , 2009, Cell cycle.

[22]  E. Slominska,et al.  1-Methylnicotinamide and nicotinamide: two related anti-inflammatory agents that differentially affect the functions of activated macrophages , 2008, Archivum Immunologiae et Therapiae Experimentalis.

[23]  C. Zheng,et al.  Correlation of Hypoxia-Inducible Factor 1α with Angiogenesis in Liver Tumors After Transcatheter Arterial Embolization in an Animal Model , 2010, CardioVascular and Interventional Radiology.

[24]  Christina Kendziorski,et al.  On Differential Variability of Expression Ratios: Improving Statistical Inference about Gene Expression Changes from Microarray Data , 2001, J. Comput. Biol..

[25]  Andrea Natali,et al.  α-Hydroxybutyrate Is an Early Biomarker of Insulin Resistance and Glucose Intolerance in a Nondiabetic Population , 2010, PloS one.

[26]  B. Efron.  Robbins, Empirical Bayes, And Microarrays , 2001 .

[27]  W. Buczko,et al.  1‐Methylnicotinamide (MNA), a primary metabolite of nicotinamide, exerts anti‐thrombotic activity mediated by a cyclooxygenase‐2/prostacyclin pathway , 2007, British journal of pharmacology.

[28]  P. Rous,et al.  EXPERIMENTS ON THE CAUSE OF THE RABBIT CARCINOMAS DERIVED FROM VIRUS-INDUCED PAPILLOMAS , 1952, The Journal of experimental medicine.

[29]  Hui Cong,et al.  Application of 1H NMR‐based metabonomics in the study of metabolic profiling of human hepatocellular carcinoma and liver cirrhosis , 2009, Cancer science.

[30]  K. Robards,et al.  Metabolomics: The greatest omics of them all? , 2006, Analytical chemistry.

[31]  M. Sherman,et al.  Screening for hepatocellular carcinoma in chronic carriers of hepatitis B virus: Incidence and prevalence of hepatocellular carcinoma in a North American urban population , 1995, Hepatology.

[32]  U. Günther,et al.  Early Stage Diagnosis of Oral Cancer Using 1 H NMR – Based Metabolomics 1,2 , 2009 .

[33]  Mark P. Styczynski,et al.  Systematic identification of conserved metabolites in GC/MS data for metabolomics and biomarker discovery. , 2007, Analytical chemistry.

[34]  Julian L. Griffin,et al.  Metabolic profiles of cancer cells , 2004, Nature Reviews Cancer.

[35]  G. Minuk.  Gaba and hepatocellular carcinoma , 2000, Molecular and Cellular Biochemistry.

[36]  Gordon K Smyth,et al.  Statistical Applications in Genetics and Molecular Biology Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2011 .

[37]  Ru Wei,et al.  The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth , 2008, Nature.

[38]  Tao-Tao Liu,et al.  A serum metabolomic investigation on hepatocellular carcinoma patients by chemical derivatization followed by gas chromatography/mass spectrometry. , 2008, Rapid communications in mass spectrometry : RCM.

[39]  J. Murai,et al.  Altered distribution and excretion of N-1-methylnicotinamide in rats with Walker 256 carcinosarcoma. , 1975, Cancer research.

[40]  Chen Huang,et al.  Gas chromatography/time-of-flight mass spectrometry-based metabonomics of hepatocarcinoma in rats with lung metastasis: elucidation of the metabolic characteristics of hepatocarcinoma at formation and metastasis. , 2010, Rapid communications in mass spectrometry : RCM.

[41]  John D. Storey.  The positive false discovery rate: a Bayesian interpretation and the q-value , 2003 .

[42]  R. Cuomo,et al.  The metabolism of nicotinamide in human liver cirrhosis: a study on N-methylnicotinamide and 2-pyridone-5-carboxamide production , 2001, American Journal of Gastroenterology.

[43]  Qing Yang,et al.  Diagnosis of liver cancer using HPLC-based metabonomics avoiding false-positive result from hepatitis and hepatocirrhosis diseases. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[44]  John D. Storey.  A direct approach to false discovery rates , 2002 .

[45]  J. Chen,et al.  Metabonomics study of liver cancer based on ultra performance liquid chromatography coupled to mass spectrometry with HILIC and RPLC separations. , 2009, Analytica chimica acta.

[46]  A. Ortega,et al.  Glucose avidity of carcinomas. , 2009, Cancer letters.

[47]  J. Kuykendall,et al.  1-Methylnicotinamide stimulates cell growth and inhibits hemoglobin synthesis in differentiating murine erythroleukemia cells. , 2007, Toxicology in vitro : an international journal published in association with BIBRA.

[48]  Peng Liu,et al.  Quick calculation for sample size while controlling false discovery rate with application to microarray analysis , 2007, Bioinform..

[49]  A. Tsirigos,et al.  Ketones and lactate “fuel” tumor growth and metastasis , 2010, Cell cycle.

[50]  W. Lin,et al.  Induction of VX2 carcinoma in rabbit liver: comparison of two inoculation methods , 2004, Laboratory animals.

[51]  R. Pankov,et al.  Ha‐ ras‐TRANSFORMATION ALTERS THE METABOLISM OF PHOSPHATIDYLETHANOLAMINE AND PHOSPHATIDYLCHOLINE IN NIH 3T3 FIBROBLASTS , 1999, Cell biology international.

[52]  Deepayan Sarkar,et al.  Detecting differential gene expression with a semiparametric hierarchical mixture method. , 2004, Biostatistics.

[53]  Andrew C Larson,et al.  Intraprocedural diffusion‐weighted PROPELLER MRI to guide percutaneous biopsy needle placement within rabbit VX2 liver tumors , 2009, Journal of magnetic resonance imaging : JMRI.

[54]  Alan Hutson,et al.  Detection of epithelial ovarian cancer using 1H‐NMR‐based metabonomics , 2005, International journal of cancer.

[55]  Manna Zhang,et al.  Increasing cell membrane potential and GABAergic activity inhibits malignant hepatocyte growth. , 2003, American journal of physiology. Gastrointestinal and liver physiology.

[56]  Peter Nilsson,et al.  Empirical Bayes Microarray ANOVA and Grouping Cell Lines by Equal Expression Levels , 2005, Statistical applications in genetics and molecular biology.

[57]  R. Shope,et al.  INFECTIOUS PAPILLOMATOSIS OF RABBITS : WITH A NOTE ON THE HISTOPATHOLOGY , 1933 .

[58]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[59]  A. Saghatelian,et al.  Exploring disease through metabolomics. , 2010, ACS chemical biology.

[60]  Hua Xu,et al.  R package MVR for Joint Adaptive Mean-Variance Regularization and Variance Stabilization. , 2011, Proceedings. American Statistical Association. Annual Meeting.

[61]  Chen Yang,et al.  Comparative Metabolomics of Breast Cancer , 2006, Pacific Symposium on Biocomputing.

[62]  Xin Lu,et al.  Integrated GC-MS and LC-MS plasma metabonomics analysis of ankylosing spondylitis. , 2008, The Analyst.