Metabolomics Analysis of the Responses to Partial Hepatectomy in Hepatocellular Carcinoma Patients

Wan Chan, Shuhai Lin, Stella Sun, Hongde Liu, John M. Luk, Zongwei Cai

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DOI: 10.4236/ajac.2011.22016                

Abstract

In this study, liquid chromatography/quadrupole time of flight mass spectrometry (LC/QTOFMS) was employed for investigating the metabolome of the sera collected from hepatocellular carcinoma (HCC) patients before and 3 to 5 months after partial hepatectomy. To investigate the changes in metabolic phenotypes after the hepatic resection, principal components analysis (PCA) and support vector machine (SVM) were performed for the data grouping and classification. Based on the obtained SVM model, mass spectrometry spectra, database searching as well as the confirmation from authentic standards, several differentiating metabolites were tentatively identified. To improve visualization, z-score plot and heat map display were performed, which exhibited the changes in concentration of the metabolites. As a result, depletion of circulating carnitine, reduced amino acid biosynthesis and increased rate of lipid peroxidation were observed. Meanwhile, up-regulation of hypoxanthine indicated that purine metabolism might serve as the salvage pathway. Collectively, the results reflected metabolic responses to surgical operation in HCC patients, suggesting perturbation of energy metabolism may occur in 3 to 5 months after the partial hepatectomy.

Keywords

Metabolomics Analysis; Partial Hepatectomy; Hepatocellular Carcinoma; LC-MS

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	R. Kaddurah-Daouk, B. S. Kristal and R. M. Weinshilboum, “Metabolomics: A Global Biochemical Approach to Drug Response and Disease,” Annual Review of Pharmacology and Toxicology, Vol. 48, 2008, pp. 653-683. doi:10.1146/annurev.pharmtox.48.113006.094715
[2]	L. M. Raamsdonk, B. Teusink, D. Broadhurst, N. Zhang, A. Hayes, M. C. Walsh, J. A. Berden, K. M. Brindle, D. B. Kell, J. J. Rowland, H. V. Westerhoff, K. Van Dam and S. G. Oliver, “A Functional Genomics Strategy That Uses Metabolome Data to Reveal the Phenotype of Silent mutations,” Nature Biotechnology, Vol. 19, No. 1, 2001, pp. 45-50.
[3]	J. R. Idle and F. J. Gonzalez, “Metabolomics,” Cell Metabolism, Vol. 6, No. 5, 2007, pp. 348-351. doi:10.1016/j.cmet.2007.10.005
[4]	J. K. Nicholson, J. C. Lindon and E. Holmes, “‘Metabonomics’: Understanding The Metabolic Responses of Living Systems to Pathophysiological Stimuli via Multivariate Statistical Analysis of Biological NMR Spectroscopic Data,” Xenobiotica, Vol. 29, No. 11, 1999, pp. 1181-1189. doi:10.1080/004982599238047
[5]	J. C. Lindon, E. Holmes, M. E. Bollard, E. G. Stanley and J. K. Nicholson, “Metabonomics Technologies and Their Applications in Physiological Monitoring, Drug Safety Assessment and Disease Diagnosis,” Biomarkers, Vol. 9, No. 1, 2004, pp. 1-31. doi:10.1080/13547500410001668379
[6]	J. L. Griffin and M. E. Bollard, “Metabonomics: Its Potential as a Tool in Toxicology for Safety Assessment and Data Integration,” Current Drug Metabolism, Vol. 5, No. 5, 2004, pp. 389-398.doi:10.2174/1389200043335432
[7]	D. G. Robertson, “Metabonomics in Toxicology: A Review,” Toxicological Sciences, Vol. 85, No. 2, 2005, pp. 809-822. doi:10.1093/toxsci/kfi102
[8]	D. B. Kell, “Metabolomics and Systems Biology: Making Sense of the Soup,” Current Opinion in Microbiology, Vol. 7, No. 3, 2004, pp. 296-307. doi:10.1016/j.mib.2004.04.012
[9]	K. Dettmer, P. A. Aronov and B. D. Hammock, “Mass Spectrometry-Based Metabolomics,” Mass Spectrometry Reviews, Vol. 26, No. 1, 2007, pp. 51-78. doi:10.1002/mas.20108
[10]	D. J. Crockford, J. C. Lindon, O. Cloarec, R. S. Plumb, S. J. Bruce, S. Zirah, P. Rainville, C. L. Stumpf, K. Johnson, E. Holmes and J. K. Nicholson, “Statistical Search Space Reduction and Two-Dimensional Data Display Approaches for UPLC-MS in Biomarker Discovery and Pathway Analysis,” Analytial Chemistry, Vol. 78, No. 13, 2006, pp. 4398-4408. doi:10.1021/ac060168o
[11]	A. Frickenschmidt, H. Frohlich, D. Bullinger, A. Zell, S. Laufer, C. H. Gleiter, H. Liebich and B. Kammerer, “Metabonomics in Cancer Diagnosis: Mass Spectrometry-Based Profiling Of Urinary Nucleosides From Breast Cancer Patients,” Biomarkers, Vol. 13, No. 4, 2008, pp. 435-449. doi:10.1080/13547500802012858
[12]	R. Xue, Z. Lin, C. Deng, L. Dong, T. Liu, J. Wang and X. Shen, “A Serum Metabolomic Investigation on Hepatocellular Carcinoma Patients by Chemical Derivatization Followed by Gas Chromatography/Mass Spectrometry,” Rapid Communications in Mass Spectrometry, Vol. 22, No. 19, 2008, pp. 3061-3068. doi:10.1002/rcm.3708
[13]	S. Mahadevan, S. L. Shah, T. J. Marrie and C. M. Slupsky, “Analysis of Metabolomic Data Using Support Vector Machines,” Analytical Chemistry, Vol. 80, No. 19, 2008, pp. 7562-7570. doi:10.1021/ac800954c
[14]	J. Yang, G. Xu, Y. Zheng, H. Kong, T. Pang, S. Lv and Q. Yang, “Diagnosis of Liver Cancer Using HPLC-Based Metabonomics Avoiding False-Positive Result from Hepatitis and Hepatocirrhosis Diseases,” Journal of Chromatography B: Analytical Technologies in Biomedical and Life Sciences, Vol. 813, No. 1-2, 2004, pp. 59-65. doi:10.1016/j.jchromb.2004.09.032
[15]	J. Yang, X. Zhao, X. Liu, C. Wang, P. Gao, J. Wang, L. Li, J. Gu, S. Yang and G. Xu, “High Performance Liquid Chromatography-Mass Spectrometry for Metabonomics: Potential Biomarkers for Acute Deterioration of Liver Function in Chronic Hepatitis B,” Journal of Proteome Research, Vol. 5, No. 3, 2006, pp. 554-561. doi:10.1021/pr050364w
[16]	K. Yu, G. Sheng, J. Sheng, Y. Chen, W. Xu, X. Liu, H. Cao, H. Qu, Y. Cheng and L. Li, “A Metabonomic Investigation on the Biochemical Perturbation in Liver Failure Patients Caused by Hepatitis B Virus,” Journal of Proteome Research, Vol. 6, No. 7, 2007, pp. 2413-2419. doi:10.1021/pr060591d
[17]	Y. Yang, C. Li, X. Nie, X. Feng, W. Chen, Y. Yue, H. Tang and F. Deng, “Metabonomic Studies of Human Hepatocellular Carcinoma Using High-Resolution Magic-Angle Spinning 1 h Nmr Spectroscopy in Conjunction with Multivariate Data Analysis,” Journal of Proteome Research, Vol. 6, No. 7, 2007, pp. 2605-2614. doi:10.1021/pr070063h
[18]	W. Chan, K. C. Lee, N. Liu, R. N. Wong, H. Liu and Z. Cai, “Liquid Chromatography/Mass Spectrometry for Metabonomics Investigation of the Biochemical Effects Induced by Aristolochic Acid in Rats: The Use of Information-Dependent Acquisition for Biomarker Identification,” Rapid Communications in Mass Spectrometry, Vol. 22, No. 6, 2008, pp. 873-880. doi:10.1002/rcm.3438
[19]	D. S. Wishart, D. Tzur, C. Knox, R. Eisner, A. C. Guo, N. Young, D. Cheng, K. Jewell, D. Arndt, S. Sawhney, C. Fung, L. Nikolai, M. Lewis, M. A. Coutouly, I. Forsythe, P. Tang, S. Shrivastava, K. Jeroncic, P. Stothard, G. Amegbey, D. Block, D. D. Hau, J. Wagner, J. Miniaci, M. Clements, M. Gebremedhin, N. Guo, Y. Zhang, G. E. Duggan, G. D. Macinnis, A. M. Weljie, R. Dowlatabadi, F. Bamforth, D. Clive, R. Greiner, L. Li, T. Marrie, B. D. Sykes, H. J. Vogel and L. Querengesser, “Hmdb: The Human Metabolome Dat
[20]	C. A. Smith, G. O’Maille, E. J. Want, C. Qin, S. A. Trauger, T. R. Brandon, D. E. Custodio, R. Abagyan and G. Siuzdak, “Metlin: A Metabolite Mass Spectral Database,” Therapeutic Drug Monitoring, Vol. 27, No. 6, 2005, pp. 747-751. doi:10.1097/01.ftd.0000179845.53213.39
[21]	S. Okuda, T. Yamada, M. Hamajima, M. Itoh, T. Katayama, P. Bork, S. Goto and M. Kanehisa, “Kegg Atlas Mapping for Global Analysis of Metabolic Pathways,” Nucleic Acids Research, Vol. 36, No. S2, 2008, pp. W423-W426.
[22]	W. R. Wikoff, G. Pendyala, G. Siuzdak and H. S. Fox, “Metabolomic Analysis of the Cerebrospinal Fluid Reveals Changes in Phospholipase Expression in the CNS of SIV-Infected Macaques,” Journal of Clinical Investigation, Vol. 118, No. 7, 2008, pp. 2661-2669. doi:10.1172/JCI34138
[23]	X. Han and R. W. Gross, “Shotgun Lipidomics: Electrospray Ionization Mass Spectrometric Analysis And Quantitation Of Cellular Lipidomes Directly From Crude Extracts Of Biological Samples,” Mass Spectrometry Reviews, Vol. 24, No. 3, 2005, pp. 367-412. doi:10.1002/mas.20023
[24]	C. Stella, B. Beckwith-Hall, O. Cloarec, E. Holmes, J. C. Lindon, J. Powell, F. Van Der Ouderaa, S. Bingham, A. J. Cross and J. K. Nicholson, “Susceptibility of Human Metabolic Phenotypes to Dietary Modulation,” Journal of Proteome Research, Vol. 5, No. 10, 2006, pp. 2780-2788. doi:10.1021/pr060265y
[25]	M. Malaguarnera, S. Restuccia, I. D. Fazio, A. M. Zoccolo, L. Ferlito and P. Bentivegna, “Serum Carnitine Levels in Chronic Hepatitis C Patients before and after Lymphoblastoid Interferon-Alpha Treatment,” BioDrugs, Vol. 12, No.1, 1999, pp. 65-69. doi:10.2165/00063030-199912010-00007
[26]	B. Chang, M. Nishikawa, S. Nishiguchi and M. Inoue, “L-Carnitine Inhibits Hepatocarcinogenesis via Protection of Mitochondria,” International Journal of Cancer, Vol. 113, No. 5, 2005, pp. 719-729. doi:10.1002/ijc.20636
[27]	P. E. Lheureux and P. Hantson, “Carnitine in the Treatment of Valproic Acid-Induced Toxicity,” Clinical Toxicology (Phila), Vol. 47, No. 2, 2009, pp. 101-111.
[28]	A. Bauer, A. Schumann, M. Gilbert, C. Wilhelm, J. G. Hengstler, J. Schiller and B. Fuchs, “Evaluation of Carbon Tetrachloride-Induced Stress on Rat Hepatocytes by 31 p Nmr And Maldi-T of Mass Spectrometry: Lysophosphatidylcholine Generation From Unsaturated Phosphatidylcholines,” Chemistry And Physics of Lipids, Vol. 159, No. 1, 2009, pp. 21-29.
[29]	P. Yin, X. Zhao, Q. Li, J. Wang, J. Li and G. Xu, “Metabonomics Study of Intestinal Fistulas Based on Ultraperformance Liquid Chromatography Coupled with Q-Tof Mass Spectrometry (UPLC/Q-TOF MS),” Journal of Proteome Research, Vol. 5, No. 9, 2006, pp. 2135-2143. doi:10.1021/pr060256p
[30]	E. S. Ong, L. Zou, S. Li, P. Y. Cheah, K. W. Eu and C. N. Ong, “Metabolic Profiling in Colorectal Cancer Reveals Signature Metabolic Shifts During Tumorigenesis,” Molecular and Cellular Proteomics, Article in press, 2010, Article ID 20147338.
[31]	V. W. Lam and R. T. Poon, “Role of Branched-Chain Amino Acids in Management of Cirrhosis and Hepatocellular Carcinoma,” Hepatology Research, Vol. 38, No. S1, 2008, pp. S107-S115. doi:10.1111/j.1872-034X.2008.00435.x
[32]	L. Vali, O. Hahn, P. Kupcsulik, A. Drahos, E. Sarvary, K. Szentmihalyi, Z. Pallai, T. Kurucz, P. Sipos and A. Blazovics, “Oxidative Stress with Altered Element Content and Decreased ATP Level of Erythrocytes in Hepatocellular Carcinoma and Colorectal Liver Metastases,” European Journal of Gastroenterology and Hepatology, Vol. 20, No. 5, 2008, pp. 393-398. doi:10.1097/MEG.0b013e3282f495c7