Hepatocellular carcinoma (HCC) is one of the most common fatal cancers worldwide. Major risk factors for HCC are chronic hepatitis resulting from infection with HBV or HCV and exposure to various exogenous carcinogens. To identify any useful biomarkers indicating virus-specific hepatocarcinogenic process, differential mRNA expression in 32 patients with HBV- / HCV-associated HCC was investigated by means of cDNA microarrays consisting of 886 genes. Among them, the up-regulation of 7 genes were involved in HBV-associated HCC, such as RPS5 (protein synthesis), KRT8 (cytoskeletal organization), CFLAR (apoptosis related genes), ATP5F1 (transport), IGFBP2 (cell membrane receptor related genes), MAP3K5 (signal transduction or transcription related genes), and MMP9 (metastasis-related genes). The up-regulated genes in HCV-infected group included 4 genes, such as VIM (cell structure), ACTB (cell structure), GAPD (glycolysis) and CD58 (cell adhesion). The different genes between HBV- and HCV-associated HCC indicate that the pathogenetic mechanisms of hepatocarcinogenesis may be different according to the viral factor, HBV and HCV. These results suggest that HBV and HCV cause hepatocarcinogenesis by different mechanisms and provide novel tools for diagnosis and treatment of HBV- and HCV- associated HCC.The high mortality associated with this disease is mainly attributed to its high tendency of metastasis. In fact, local lymph node and blood metastasis could occur at early stage, which may be the key factors of its recurrence and poor prognosis. Therefore, a better understanding for the molecular mechanism of metastasis can improve prevention and treatment of HCC. Our results of the cDNA microarray demonstrated that 25 up-regulated genes that included cell adhesion related genes (CD58, ITGAM, ITGB5, OPCML), cell membrane receptor (CD86, JUN, MET, EPHA2, EGFR), cell death regulator (BNIP3, SEMA4D), intracellular signaling related genes (ARHGDIG, RAC3, IGFBP3, F2R), growth/differentiation factor (VEGFC), cell cycle regulator (CCND1, CDK4), and other genes (TIMP3, UBE2A, YES1, POR, MMP13, MMP2). The 48 down-regulated genes included cell adhesion related genes (DSP, PCDHGC3, ITGB4, ITGA3, CTNNA1, CDH1), cell cycle regulator (CDC25B, CDK5, CCND2, MAPRE1, PPM1D, ATR, PPP2R5A), intracellular signaling related genes (MAPK14, MAPK7, MAPK4, SCYB13, STAT5B, SERPINB1, SERPINB2, SH3BP2, GPRK6, RAGB, PTPRF), cell membrane receptor (EPHB6, KAI1, SCYA25, MGLL, IFNAR1, IGFBP6), metabolic enzyme (SPINT2, PPP2R5A, PTPN2, DNASE1L3, CTSL), cell death regulator (CASP3, PDCD10), DNA damage response related genes (ATR, XRCC5, MDM2), growth/differentiation factor (BMP5, KRT4, KRT13, CTGF), and other genes (HSPA4, RXRA, UBE1L, SLC25A5).
The result of this study may help clinicians to predict metastasis of lymph nodes and assist researchers in identifying novel therapeutic targets for metastatic HCC patients. However, it is more important to further analyze each gene in detail to elucidate the mechanism of lymph node metastasis in HCC.

Aishima S, Nishihara Y, Iguchi T, Taguchi K, Taketomi A, Maehara Y, Tsuneyoshi M. Lymphatic spread is related to VEGF-C expression and D2-40-positive myofibroblasts in intrahepatic cholangiocarcinoma. Mod Pathol. 2008 Mar;21(3):256-64.

Alizadeh, A.A., et al. Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature 2000; 403,503–511

Beasley RP, Hwang LY, Lin CC, Chien CS. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22707 men in Taiwan. Lancet 1981;21:1129-33.

Bowen, D. G. & Walker, C. M. Adaptive immune responses in acute and chronic hepatitis C virus infection. 2005 Nature 436, 946-952

Delpuech O, Trabut JB, Carnot F, Feuillard J, Brechot C, Kremsdorf D. Identification, using cDNA macroarray analysis, of distinct gene expression profiles associated with pathological and virological features of hepatocellular carcinoma. Oncogene 2002; 21: 2926-2937

Deuffic S, Poynard T, Buffat L,Valleron AJ. Trends in primary liver cancer. Lancet 1998;351:214-5.

El-Serag, H. B., Mason A. C. Rising incidence of hepatocellular carcinoma in the United States. N. Engl. J. Med 1999; 340: 745–750

Floris G, Debiec-Rychter M, Wozniak A, Magrini E, Manfioletti G, De Wever I, Tallini G, Sciot R. Malignant ectomesenchymoma: genetic profile reflects rhabdomyosarcomatous differentiation. Diagn Mol Pathol. 2007 Dec;16(4):243-8.

Gazitt Y. Homing and mobilization of hematopoietic stem cells and hematopoietic cancer cells are mirror image processes, utilizing similar signaling pathways and occurring concurrently: circulating cancer cells constitute an ideal target for concurrent treatment with chemotherapy and antilineage-specific antibodies. Leukemia. 2004 Jan;18(1):1-10.

Grant SF, Hakonarson H. Microarray technology and applications in the arena of genome-wide association. Clin Chem. 2008 Jul;54(7):1116-24. Epub 2008 May 22.

Gruber HE, Mougeot JL, Hoelscher G, Ingram JA, Hanley EN Jr. Microarray analysis of laser capture microdissected-anulus cells from the human intervertebral disc. Spine. 2007 May 15;32(11):1181-7.

Gu T, Kilinc MO, Egilmez NK. Transient activation of tumor-associated T-effector/memory cells promotes tumor eradication via NK-cell recruitment: minimal role for long-term T-cell immunity in cure of metastatic disease. Cancer Immunol Immunother. 2008 Jul;57(7):997-1005.

Hirakawa S, Hong YK, Harvey N, Schacht V, Matsuda K, Libermann T, Detmar M. Identification of vascular lineage-specific genes by transcriptional profiling of isolated blood vascular and lymphatic endothelial cells. Am J Pathol. 2003 Feb;162(2):575-86.

Hoek, J. B. & Pastorino, J. G. Ethanol oxidative stress, and cytokine-induced liver cell injury. 2002 Alcohol 27, 63-68

Honda, M., Kaneko, S., Kawai, H., Shirota, Y., and Kobayash, I. K. Differential gene expression between chronic hepatitis b and c hepatic lesion. Gastroenterology 2000; 120:955–966

Iizuka N, Hamamoto Y, Tsunedomi R, Oka M. Translational microarray systems for outcome prediction of hepatocellular carcinoma. Cancer Sci. 2008 Apr;99(4):659-65.

Julien SG, Dub□ N, Read M, Penney J, Paquet M, Han Y, Kennedy BP, Muller WJ, Tremblay ML. Protein tyrosine phosphatase 1B deficiency or inhibition delays ErbB2-induced mammary tumorigenesis and protects from lung metastasis. Nat Genet. 2007 Mar;39(3):338-46.

Kasai Y, Takeda S and Takagi H. Pathogenesis of hepatocellular carcinoma: a review from the viewpoint of molecular analysis. Semin Surg Oncol 1996; 12: 155-159

Kerbauy DB, Deeg HJ. Apoptosis and antiapoptotic mechanisms in the progression of myelodysplastic syndrome. Exp Hematol. 2007 Nov;35(11):1739

Kishimoto I, Mitomi H, Ohkura Y, Kanazawa H, Fukui N, Watanabe M. Abnormal expression of p16(INK4a), cyclin D1, cyclin-dependent kinase 4 and retinoblastoma protein in gastric carcinomas. J Surg Oncol. 2008 Jul 1; 98(1):60-66.

Kovacevic Z, Richardson DR. The metastasis suppressor, Ndrg-1: a new ally in the fight against cancer. Carcinogenesis. 2006 Dec;27(12):2355-66.

K□bel M, Huntsman D, Gilks CB. Critical molecular abnormalities in high-grade serous carcinoma of the ovary. Expert Rev Mol Med. 2008 Aug 1;10:e22.

Kuehn H, Liberzon A, Reich M, Mesirov JP. Using GenePattern for gene expression analysis. Curr Protoc Bioinformatics. 2008 Jun;Chapter 7:Unit 7.12.

Lavanchy, D. Hepatitis B epidemiology, disease burden, treatment, and current and emerging prevention and control measure. 2004 J. Viral. Hepat. 11, 97-107

Lee, W. N. Hepatitis B virus infection. N. Engl. J. Med 1997; 337: 1733–1745

Le Bail B, Faouzi S, Boussarie L, Balabaud C, Bioulac-Sage P, Rosenbaum J. Extracellular matrix composition and integrin expression in early hepatocarcinogenesis in human cirrhotic liver. J Pathol 1997;181: 330-337

Liu B, Yu J, Taylor L, Zhou X, Polgar P. Microarray and phosphokinase screenings leading to studies on ERK and JNK regulation of connective tissue growth factor expression by angiotensin II 1a and bradykinin B2 receptors in Rat1 fibroblasts. J Cell Biochem. 2006 Apr 1;97(5):1104-20.
Lunt SJ, Chaudary N, Hill RP. The tumor microenvironment and metastatic disease. Clin Exp Metastasis. 2008 Jun 10. [Epub ahead of print]

Li Y, Tian B, Yang J, Zhao L, Wu X, Ye SL, Liu YK, Tang ZY. Stepwise metastatic human hepatocellular carcinoma cell model system with multiple metastatic potentials established through consecutive in vivo selection and studies on metastatic characteristics. J Cancer Res Clin Oncol. 2004 Aug;130(8):460-8.

Li T, Ling YH, Perez-Soler R. Tumor dependence on the EGFR signaling pathway expressed by the p-EGFR:p-AKT ratio predicts erlotinib sensitivity in human non-small cell lung cancer (NSCLC) cells expressing wild-type EGFR gene. J Thorac Oncol. 2008 Jun;3(6):643-7.

Liu S, Yu M, He Y, Xiao L, Wang F, Song C, Sun S, Ling C, Xu Z. Melittin prevents liver cancer cell metastasis through inhibition of the Rac1-dependent pathway. Hepatology. 2008 Jun;47(6):1964-73.

Mishra PJ, Mishra PJ, Humeniuk R, Medina DJ, Alexe G, Mesirov JP, Ganesan S, Glod JW, Banerjee D. Carcinoma-associated fibroblast-like differentiation of human mesenchymal stem cells. Cancer Res. 2008 Jun 1;68(11):4331-9.

Mudali SV, Fu B, Lakkur SS, Luo M, Embuscado EE, Iacobuzio-Donahue CA. Patterns of EphA2 protein expression in primary and metastatic pancreatic carcinoma and correlation with genetic status. Clin Exp Metastasis. 2006;23(7-8):357-65.

Nakamoto M, Bergemann AD. Diverse roles for the Eph family of receptor tyrosine kinases in carcinogenesis. Microsc Res Tech. 2002 Oct 1;59(1):58-67.

N. Rino Y, Masuda M, Tanaka K, Imada T. Clinicopathological significance of the gene expression of matrix metalloproteinases and reversion-inducing cysteine-rich protein with Kazal motifs in patients with colorectal cancer: MMP-2 gene expression is a useful predictor of liver metastasis from colorectal cancer. Oncol Rep. 2008 May;19(5):1285-91.

Okabe, H., Satoh, S., Kato, T., Kitahara, O., Yanagawa, R., Yamaoka, Y., Tsunoda, T., Furukawa, Y., Nakamura Y. Genome-wide analysis of gene expression in human hepatocellular carcinomas using cDNA microarray: identification of genes involved in viral carcinogenesis and tumor progression. Cancer Res 2001; 61: 2129–2137

Okuda K, Fujimoto I, Hanai A and Urano Y. Changing incidence of hepatocellular carcinoma in Japan. Cancer Res 1987; 47:4967-4972

Oshima T, Kunisaki C, Yoshihara K, Yamada R, Yamamoto N, Sato T, Makino H, Yamagishi S, Nagano Y, Fujii S, Shiozawa M, Akaike M, Wada

Parkin DM, Pisani P and Ferlay J. Global cancer statistics. CA Cancer J Clin 1999; 49: 33-64

Pang H, Rowan BG, Al-Dhaheri M, Faber LE. Epidermal growth factor suppresses induction by progestin of the adhesion protein desmoplakin in T47D breast cancer cells. Breast Cancer Res. 2004;6(3):R239-45.

Perou, C.M., et al. Distinctive gene expression patterns in human mammary epithelial cells and breast cancers. Proc. Natl. Acad. Sci. USA1999; 96, 9212–9217

Perou, C.M., et al. Molecular portraits of human breast tumors. Nature 2000; 406, 747–752

Protzel C, Kakies C, Kleist B, Poetsch M, Giebel J. Down-regulation of the metastasis suppressor protein KAI1/CD82 correlates with occurrence of metastasis, prognosis and presence of HPV DNA in human penile squamous cell carcinoma. Virchows Arch. 2008 Apr;452(4):369-75.

Qu Z, Van Ginkel S, Roy AM, Westbrook L, Nasrin M, Maxuitenko Y, Frost AR, Carey D, Wang W, Li R, Grizzle WE, Thottassery JV, Kern FG. Vascular endothelial growth factor reduces tamoxifen efficacy and promotes metastatic colonization and desmoplasia in breast tumors. Cancer Res. 2008 Aug 1;68(15):6232-40.

Robinson DR, Zylstra CR, Williams BO. Wnt signaling and prostate cancer. Curr Drug Targets. 2008 Jul;9(7):571-80.

Ross, D.T., et al. Systematic variation in gene expression patterns in human cancer cell lines. Nat. Genet 2000; 24, 227–235

Rydlova M, Holubec L Jr, Ludvikova M Jr, Kalfert D, Franekova J, Povysil C, Ludvikova M. Biological activity and clinical implications of the matrix metalloproteinases. Anticancer Res. 2008 Mar-Apr;28(2B):1389-97.

Riker AI, Enkemann SA, Fodstad O, Liu S, Ren S, Morris C, Xi Y, Howell P, Metge B, Samant RS, Shevde LA, Li W, Eschrich S, Daud A, Ju J, Matta J. The gene expression profiles of primary and metastatic melanoma yields a transition point of tumor progression and metastasis. BMC Med Genomics. 2008 Apr 28;1:13.

Rizki A, Weaver VM, Lee SY, Rozenberg GI, Chin K, Myers CA, Bascom JL, Mott JD, Semeiks JR, Grate LR, Mian IS, Borowsky AD, Jensen RA, Idowu MO, Chen F, Chen DJ, Petersen OW, Gray JW, Bissell MJ. A human breast cell model of preinvasive to invasive transition. Cancer Res. 2008 Mar 1;68(5):1378-87.

Sanchez G, Delattre O, Auboeuf D, Dutertre M. Coupled alteration of transcription and splicing by a single oncogene: boosting the effect on cyclin D1 activity. Cell Cycle. 2008 Aug;7(15):2299-305.

Sabates-Bellver J, Van der Flier LG, de Palo M, Cattaneo E, Maake C, Rehrauer H, Laczko E, Kurowski MA, Bujnicki JM, Menigatti M, Luz J, Ranalli TV, Gomes V, Pastorelli A, Faggiani R, Anti M, Jiricny J, Clevers H, Marra G. Transcriptome profile of human colorectal adenomas. Mol Cancer Res. 2007 Dec;5(12):1263-75.

Sampaio C, Dance M, Montagner A, Edouard T, Malet N, Perret B, Yart A, Salles JP, Raynal P. Signal strength dictates phosphoinositide 3-kinase contribution to Ras/extracellular signal-regulated kinase 1 and 2 activation via differential Gab1/Shp2 recruitment: consequences for resistance to epidermal growth factor receptor inhibition. Mol Cell Biol. 2008 Jan;28(2):587-600.

Strong VE, Kennedy T, Al-Ahmadie H, Tang L, Coleman J, Fong Y, Brennan M, Ghossein RA. Prognostic indicators of malignancy in adrenal pheochromocytomas: clinical, histopathologic, and cell cycle/apoptosis gene expression analysis. Surgery. 2008 Jun;143(6):759-68.

Tachibana M, Tonomoto Y, Hyakudomi R, Hyakudomi M, Hattori S, Ueda S, Kinugasa S, Yoshimura H. Expression and prognostic significance of EFNB2 and EphB4 genes in patients with oesophageal squamous cell carcinoma. Dig Liver Dis. 2007 Aug;39(8):725-32.

Tanaka N, Odajima T, Ogi K, Ikeda T, Satoh M. Expression of E-cadherin, alpha-catenin, and beta-catenin in the process of lymph node metastasis in oral squamous cell carcinoma. Br J Cancer. 2003 Aug 4;89(3):557-63.

Taylor-Robinson SD, Foster GR, Arora S, Hargreaves S,Thomas HC. Increase in primary liver cancer in the UK, 1979-94. Lancet 1997;350:1142-3.

Turner DP, Findlay VJ, Kirven AD, Moussa O, Watson DK. Global Gene Expression Analysis Identifies PDEF Transcriptional Networks Regulating Cell Migration during Cancer Progression. Mol Biol Cell. 2008 Jun 25. [Epub ahead of print].

Waddell N. Microarray-based DNA profiling to study genomic aberrations. IUBMB Life. 2008 Jul;60(7):437-40.

Welsh, J.B., Zarrinkar, P.P., Sapinoso, L.M., Kern, S.G., Behling, C.A., Monk, B.J., Lockhart, D.J., Burger, R.A., and Hampton, G.M. Analysis of gene expression profiles in normal and neoplastic ovarian tissue samples identifies candidate molecular markers of epithelial ovarian cancer. Proc. Natl. Acad. Sci. USA 2001; 98, 1176–1181

Wu Y, Zhou BP. New insights of epithelial-mesenchymal transition in cancer metastasis. Acta Biochim Biophys Sin (Shanghai). 2008 Jul; 40(7):643-50.

Wu CG, Forgues M, Siddique S, Farnsworth J, Valerie K, Wang XW. SAGE transcript profiles of normal primary human hepatocytes expressing oncogenic hepatitis B virus X protein. FASEB J 2002;16: 1665-1667

Xu, XR, Huang J, Xu ZG, Qian BZ, Zhu ZD, Yan Q, Cai T, Zhang X, Xiao HS, Qu J, Liu F, Huang QH, Cheng ZH, Li NG, Du JJ, Hu W, Shen KT, Lu G, Fu G, Zhong M, Xu SH, Gu WY, Huang W, Zhao XT, Hu G.X, Gu JR, Chen Z, Han ZG. Insight into hepatocellular carcinogenesis at transcriptome level by comparing gene expression profiles of hepatocellular carcinoma with those of corresponding noncancerous liver. Proc Natl Acad Sci USA 2001; 98: 15089–15094

Yan B, Chen G, Saigal K, Yang X, Jensen ST, Van Waes C, Stoeckert CJ, Chen Z. Systems biology-defined NF-kappaB regulons, interacting signal pathways and networks are implicated in the malignant phenotype of head and neck cancer cell lines differing in p53 status. Genome Biol. 2008;9(3):R53. Epub 2008 Mar 11.

Yoon SY, Kim JM, Oh JH, Jeon YJ, Lee DS, Kim JH, Choi JY, Ahn BM, Kim SS, Yoo HS, Kim YS, Kim NS. Gene expression profiling of human HBV- and/or HCV-associated hepatocellular carcinoma cells using expressed sequence tags. Int J Oncol, 2006;29:315-327

Yu J, Becka S, Zhang P, Zhang X, Brady-Kalnay SM, Wang Z. Tumor-derived extracellular mutations of PTPRT /PTPrho are defective in cell adhesion. Mol Cancer Res. 2008 Jul;6(7):1106-13.

Ziegler DS, Kung AL, Kieran MW. Anti-apoptosis mechanisms in malignant gliomas. J Clin Oncol. 2008 Jan 20;26(3):493-500.

Zhao P, Lu Y, Zhong M, Liu L, Li B. Inverse correlation of aberrant expression of fragile histidine triad (FHIT) protein with cyclin D1 protein and prognosis in Chinese patients with cholangiocarcinoma. Acta Oncol 2008; 47:1557-1563

Zhuang PY, Zhang JB, Zhu XD, Zhang W, Wu WZ, Tan YS, Hou J, Tang ZY, Qin LX, Sun HC. Two pathologic types of hepatocellular carcinoma with lymph node metastasis with distinct prognosis on the basis of CK19 expression in tumor. Cancer. 2008 Jun 15;112(12):2740-8.