Expression Profiling of Hepatocellular Carcinoma

  • Rosina Maria Critelli
  • Elisabetta Cariani
  • Erica Villa
Chapter
First Online:

Abstract

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths in the world.

The molecular mechanism of HCC onset involves a complex interplay of both genetic and epigenetic factors. Hepatic carcinogenesis is characterized by an increase in allelic losses, chromosomal aberrations, gene mutations, epigenetic alterations, changes of gene expression and alterations in molecular cellular pathways. The integration of genetic, epigenetic, genomic, and proteomic data provides insight into the molecular mechanisms underlying hepatocarcinogenesis and is revealing promising clinical approaches. Resulting findings offer the possibility for the identification of relevant biomarkers, useful for the detection, molecular diagnosis, prediction of recurrence and prognosis of HCC as well as for the improved identification of novel therapeutic targets. This will be of utmost importance in the near future as more and more new targeted drugs will become available.

Keywords

Hepatocellular carcinoma mRNA profiling Genomic profiling miRNAs Methylation Histone modification 
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References

  1. 1.
    El-Serag HB, Rudolph KL (2007) Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology 132(7):2557–2576. doi: S0016-5085(07)00799-8[pii]10.1053/j.gastro.2007.04.061PubMedCrossRefGoogle Scholar
  2. 2.
    Llovet JM, Brú C, Bruix J (1999) Prognosis of hepatocellular carcinoma: the BCLC staging classification. Semin Liver Dis 19(3):329–338. doi: 10.1055/s-2007-1007122PubMedCrossRefGoogle Scholar
  3. 3.
    Villanueva A, Newell P, Chiang DY, Friedman SL, Llovet JM (2007) Genomics and signaling pathways in hepatocellular carcinoma. Semin Liver Dis 27(1):55–76. doi: 10.1055/s-2006-960171PubMedCrossRefGoogle Scholar
  4. 4.
    Kaposi-Novak P, Lee JS, Gòmez-Quiroz L, Coulouarn C, Factor VM, Thorgeirsson SS (2006) Met-regulated expression signature defines a subset of human hepatocellular carcinomas with poor prognosis and aggressive phenotype. J Clin Invest 116(6):1582–1595. doi: 10.1172/JCI27236PubMedCrossRefGoogle Scholar
  5. 5.
    Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, Schwartz M, Porta C, Zeuzem S, Bolondi L, Greten TF, Galle PR, Seitz JF, Borbath I, Häussinger D, Giannaris T, Shan M, Moscovici M, Voliotis D, Bruix J, Group SIS (2008) Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 359(4):378–390. doi: 359/4/378[pii]10.1056/NEJMoa0708857PubMedCrossRefGoogle Scholar
  6. 6.
    Finn RS (2010) Development of molecularly targeted therapies in hepatocellular carcinoma: where do we go now? Clin Cancer Res 16(2):390–397. doi: 1078-0432.CCR-09-2084[pii]10.1158/1078-0432.CCR-09-2084PubMedCrossRefGoogle Scholar
  7. 7.
    Chiang DY, Villanueva A, Hoshida Y, Peix J, Newell P, Minguez B, LeBlanc AC, Donovan DJ, Thung SN, Solé M, Tovar V, Alsinet C, Ramos AH, Barretina J, Roayaie S, Schwartz M, Waxman S, Bruix J, Mazzaferro V, Ligon AH, Najfeld V, Friedman SL, Sellers WR, Meyerson M, Llovet JM (2008) Focal gains of VEGFA and molecular classification of hepatocellular carcinoma. Cancer Res 68(16):6779–6788. doi: 68/16/6779[pii]10.1158/0008-5472.CAN-08-0742PubMedCrossRefGoogle Scholar
  8. 8.
    Hoshida Y, Nijman SM, Kobayashi M, Chan JA, Brunet JP, Chiang DY, Villanueva A, Newell P, Ikeda K, Hashimoto M, Watanabe G, Gabriel S, Friedman SL, Kumada H, Llovet JM, Golub TR (2009) Integrative transcriptome analysis reveals common molecular subclasses of human hepatocellular carcinoma. Cancer Res 69(18):7385–7392. doi: 0008-5472.CAN-09-1089[pii]10.1158/0008-5472.CAN-09-1089PubMedCrossRefGoogle Scholar
  9. 9.
    Lee JS, Chu IS, Heo J, Calvisi DF, Sun Z, Roskams T, Durnez A, Demetris AJ, Thorgeirsson SS (2004) Classification and prediction of survival in hepatocellular carcinoma by gene expression profiling. Hepatology 40(3):667–676. doi: 10.1002/hep.20375PubMedCrossRefGoogle Scholar
  10. 10.
    Lee JS, Heo J, Libbrecht L, Chu IS, Kaposi-Novak P, Calvisi DF, Mikaelyan A, Roberts LR, Demetris AJ, Sun Z, Nevens F, Roskams T, Thorgeirsson SS (2006) A novel prognostic subtype of human hepatocellular carcinoma derived from hepatic progenitor cells. Nat Med 12(4):410–416. doi: nm1377[pii]10.1038/nm1377PubMedCrossRefGoogle Scholar
  11. 11.
    Breuhahn K, Vreden S, Haddad R, Beckebaum S, Stippel D, Flemming P, Nussbaum T, Caselmann WH, Haab BB, Schirmacher P (2004) Molecular profiling of human hepatocellular carcinoma defines mutually exclusive interferon regulation and insulin-like growth factor II overexpression. Cancer Res 64(17):6058–6064. doi: 64/17/6058[pii]10.1158/0008-5472.CAN-04-0292PubMedCrossRefGoogle Scholar
  12. 12.
    Boyault S, Rickman DS, de Reyniès A, Balabaud C, Rebouissou S, Jeannot E, Hérault A, Saric J, Belghiti J, Franco D, Bioulac-Sage P, Laurent-Puig P, Zucman-Rossi J (2007) Transcriptome classification of HCC is related to gene alterations and to new therapeutic targets. Hepatology 45(1):42–52. doi: 10.1002/hep.21467PubMedCrossRefGoogle Scholar
  13. 13.
    Yamashita T, Forgues M, Wang W, Kim JW, Ye Q, Jia H, Budhu A, Zanetti KA, Chen Y, Qin LX, Tang ZY, Wang XW (2008) EpCAM and alpha-fetoprotein expression defines novel prognostic subtypes of hepatocellular carcinoma. Cancer Res 68(5):1451–1461. doi: 68/5/1451[pii]10.1158/0008-5472.CAN-07-6013PubMedCrossRefGoogle Scholar
  14. 14.
    Yang JD, Nakamura I, Roberts LR (2011) The tumor microenvironment in hepatocellular carcinoma: current status and therapeutic targets. Semin Cancer Biol 21(1):35–43. doi: S1044-579X(10)00091-X[pii]10.1016/j.semcancer.2010.10.007PubMedCrossRefGoogle Scholar
  15. 15.
    Fu J, Xu D, Liu Z, Shi M, Zhao P, Fu B, Zhang Z, Yang H, Zhang H, Zhou C, Yao J, Jin L, Wang H, Yang Y, Fu YX, Wang FS (2007) Increased regulatory T cells correlate with CD8 T-cell impairment and poor survival in hepatocellular carcinoma patients. Gastroenterology 132(7):2328–2339. doi: S0016-5085(07)00628-2[pii]10.1053/j.gastro.2007.03.102PubMedCrossRefGoogle Scholar
  16. 16.
    Kobayashi N, Hiraoka N, Yamagami W, Ojima H, Kanai Y, Kosuge T, Nakajima A, Hirohashi S (2007) FOXP3+ regulatory T cells affect the development and progression of hepatocarcinogenesis. Clin Cancer Res 13(3):902–911. doi: 13/3/902[pii]10.1158/1078-0432.CCR-06-2363PubMedCrossRefGoogle Scholar
  17. 17.
    Gao Q, Qiu SJ, Fan J, Zhou J, Wang XY, Xiao YS, Xu Y, Li YW, Tang ZY (2007) Intratumoral balance of regulatory and cytotoxic T cells is associated with prognosis of hepatocellular carcinoma after resection. J Clin Oncol 25(18):2586–2593. doi: 25/18/2586[pii]10.1200/JCO.2006.09.4565PubMedCrossRefGoogle Scholar
  18. 18.
    Chew V, Tow C, Teo M, Wong HL, Chan J, Gehring A, Loh M, Bolze A, Quek R, Lee VK, Lee KH, Abastado JP, Toh HC, Nardin A (2010) Inflammatory tumour microenvironment is associated with superior survival in hepatocellular carcinoma patients. J Hepatol 52(3):370–379. doi: S0168-8278(09)00475-9[pii]10.1016/j.jhep.2009.07.013PubMedCrossRefGoogle Scholar
  19. 19.
    Dong H, Strome SE, Salomao DR, Tamura H, Hirano F, Flies DB, Roche PC, Lu J, Zhu G, Tamada K, Lennon VA, Celis E, Chen L (2002) Tumor-associated B7-H1 promotes T-cell apoptosis: a potential mechanism of immune evasion. Nat Med 8(8):793–800. doi: nm730[pii]10.1038/nm730PubMedGoogle Scholar
  20. 20.
    Gao Q, Wang XY, Qiu SJ, Yamato I, Sho M, Nakajima Y, Zhou J, Li BZ, Shi YH, Xiao YS, Xu Y, Fan J (2009) Overexpression of PD-L1 significantly associates with tumor aggressiveness and postoperative recurrence in human hepatocellular carcinoma. Clin Cancer Res 15(3):971–979. doi: 15/3/971[pii]10.1158/1078-0432.CCR-08-1608PubMedCrossRefGoogle Scholar
  21. 21.
    Kuang DM, Zhao Q, Peng C, Xu J, Zhang JP, Wu C, Zheng L (2009) Activated monocytes in peritumoral stroma of hepatocellular carcinoma foster immune privilege and disease progression through PD-L1. J Exp Med 206(6):1327–1337. doi: jem.20082173[pii]10.1084/jem.20082173PubMedCrossRefGoogle Scholar
  22. 22.
    Shi F, Shi M, Zeng Z, Qi RZ, Liu ZW, Zhang JY, Yang YP, Tien P, Wang FS (2011) PD-1 and PD-L1 upregulation promotes CD8(+) T-cell apoptosis and postoperative recurrence in hepatocellular carcinoma patients. Int J Cancer 128(4):887–896. doi: 10.1002/ijc.25397PubMedCrossRefGoogle Scholar
  23. 23.
    Budhu A, Forgues M, Ye QH, Jia HL, He P, Zanetti KA, Kammula US, Chen Y, Qin LX, Tang ZY, Wang XW (2006) Prediction of venous metastases, recurrence, and prognosis in hepatocellular carcinoma based on a unique immune response signature of the liver microenvironment. Cancer Cell 10(2):99–111. doi: S1535-6108(06)00214-5[pii]10.1016/j.ccr.2006.06.016PubMedCrossRefGoogle Scholar
  24. 24.
    Hoshida Y, Villanueva A, Kobayashi M, Peix J, Chiang DY, Camargo A, Gupta S, Moore J, Wrobel MJ, Lerner J, Reich M, Chan JA, Glickman JN, Ikeda K, Hashimoto M, Watanabe G, Daidone MG, Roayaie S, Schwartz M, Thung S, Salvesen HB, Gabriel S, Mazzaferro V, Bruix J, Friedman SL, Kumada H, Llovet JM, Golub TR (2008) Gene expression in fixed tissues and outcome in hepatocellular carcinoma. N Engl J Med 359(19):1995–2004. doi: NEJMoa0804525[pii]10.1056/NEJMoa0804525PubMedCrossRefGoogle Scholar
  25. 25.
    Sherman M (2008) Recurrence of hepatocellular carcinoma. N Engl J Med 359(19):2045–2047. doi: NEJMe0807581[pii]10.1056/NEJMe0807581PubMedCrossRefGoogle Scholar
  26. 26.
    Tsuchiya M, Parker JS, Kono H, Matsuda M, Fujii H, Rusyn I (2010) Gene expression in nontumoral liver tissue and recurrence-free survival in hepatitis C virus-positive hepatocellular carcinoma. Mol Cancer 9:74. doi: 1476-4598-9-74[pii]10.1186/1476-4598-9-74PubMedCrossRefGoogle Scholar
  27. 27.
    Tanaka S, Mogushi K, Yasen M, Ban D, Noguchi N, Irie T, Kudo A, Nakamura N, Tanaka H, Yamamoto M, Kokudo N, Takayama T, Kawasaki S, Sakamoto M, Arii S (2011) Oxidative stress pathways in noncancerous human liver tissue to predict hepatocellular carcinoma recurrence: a prospective, multicenter study. Hepatology 54(4):1273–1281. doi: 10.1002/hep.24536PubMedCrossRefGoogle Scholar
  28. 28.
    Hoshida Y, Toffanin S, Lachenmayer A, Villanueva A, Minguez B, Llovet JM (2010) Molecular classification and novel targets in hepatocellular carcinoma: recent advancements. Semin Liver Dis 30(1):35–51. doi: 10.1055/s-0030-1247131PubMedCrossRefGoogle Scholar
  29. 29.
    Villanueva A, Hoshida Y, Battiston C, Tovar V, Sia D, Alsinet C, Cornella H, Liberzon A, Kobayashi M, Kumada H, Thung SN, Bruix J, Newell P, April C, Fan JB, Roayaie S, Mazzaferro V, Schwartz ME, Llovet JM (2011) Combining clinical, pathology, and gene expression data to predict recurrence of hepatocellular carcinoma. Gastroenterology 140(5):1501–1512. doi: S0016-5085(11)00142-9[pii]10.1053/j.gastro.2011.02.006, e1502PubMedCrossRefGoogle Scholar
  30. 30.
    Kallioniemi A, Kallioniemi OP, Sudar D, Rutovitz D, Gray JW, Waldman F, Pinkel D (1992) Comparative genomic hybridization for molecular cytogenetic analysis of solid tumors. Science 258(5083):818–821PubMedCrossRefGoogle Scholar
  31. 31.
    Thorgeirsson SS, Grisham JW (2002) Molecular pathogenesis of human hepatocellular carcinoma. Nat Genet 31(4):339–346. doi: ng0802-339[pii]10.1038/ng0802-339PubMedCrossRefGoogle Scholar
  32. 32.
    Homayounfar K, Gunawan B, Cameron S, Haller F, Baumhoer D, Uecker S, Sander B, Ramadori G, Lorf T, Füzesi L (2009) Pattern of chromosomal aberrations in primary liver cancers identified by comparative genomic hybridization. Hum Pathol 40(6):834–842. doi: S0046-8177(08)00560-1[pii]10.1016/j.humpath.2008.11.005PubMedCrossRefGoogle Scholar
  33. 33.
    Moinzadeh P, Breuhahn K, Stützer H, Schirmacher P (2005) Chromosome alterations in human hepatocellular carcinomas correlate with aetiology and histological grade–results of an explorative CGH meta-analysis. Br J Cancer 92(5):935–941. doi: 6602448[pii]10.1038/sj.bjc.6602448PubMedCrossRefGoogle Scholar
  34. 34.
    Poon TC, Wong N, Lai PB, Rattray M, Johnson PJ, Sung JJ (2006) A tumor progression model for hepatocellular carcinoma: bioinformatic analysis of genomic data. Gastroenterology 131(4):1262–1270. doi: S0016-5085(06)01753-7[pii]10.1053/j.gastro.2006.08.014PubMedCrossRefGoogle Scholar
  35. 35.
    Laurent-Puig P, Legoix P, Bluteau O, Belghiti J, Franco D, Binot F, Monges G, Thomas G, Bioulac-Sage P, Zucman-Rossi J (2001) Genetic alterations associated with hepatocellular carcinomas define distinct pathways of hepatocarcinogenesis. Gastroenterology 120(7):1763–1773. doi: S0016508501008228[pii]PubMedCrossRefGoogle Scholar
  36. 36.
    Nagai H, Pineau P, Tiollais P, Buendia MA, Dejean A (1997) Comprehensive allelotyping of human hepatocellular carcinoma. Oncogene 14(24):2927–2933. doi: 10.1038/sj.onc.1201136PubMedCrossRefGoogle Scholar
  37. 37.
    Wong CM, Fan ST (2001) Ng, I.O.: beta-Catenin mutation and overexpression in hepatocellular carcinoma: clinicopathologic and prognostic significance. Cancer 92(1):136–145. doi:10.1002/1097-0142(20010701)92:1<136::AID-CNCR1301>3.0.CO;2-R [pii]PubMedCrossRefGoogle Scholar
  38. 38.
    Schlaeger C, Longerich T, Schiller C, Bewerunge P, Mehrabi A, Toedt G, Kleeff J, Ehemann V, Eils R, Lichter P, Schirmacher P, Radlwimmer B (2008) Etiology-dependent molecular mechanisms in human hepatocarcinogenesis. Hepatology 47(2):511–520. doi: 10.1002/hep.22033PubMedCrossRefGoogle Scholar
  39. 39.
    Katoh H, Shibata T, Kokubu A, Ojima H, Loukopoulos P, Kanai Y, Kosuge T, Fukayama M, Kondo T, Sakamoto M, Hosoda F, Ohki M, Imoto I, Inazawa J, Hirohashi S (2005) Genetic profile of hepatocellular carcinoma revealed by array-based comparative genomic hybridization: identification of genetic indicators to predict patient outcome. J Hepatol 43(5):863–874. doi: S0168-8278(05)00415-0[pii]10.1016/j.jhep.2005.05.033PubMedCrossRefGoogle Scholar
  40. 40.
    Bressac B, Kew M, Wands J, Ozturk M (1991) Selective G to T mutations of p53 gene in hepatocellular carcinoma from southern Africa. Nature 350(6317):429–431. doi: 10.1038/350429a0PubMedCrossRefGoogle Scholar
  41. 41.
    Clifford RJ, Zhang J, Meerzaman DM, Lyu MS, Hu Y, Cultraro CM, Finney RP, Kelley JM, Efroni S, Greenblum SI, Nguyen CV, Rowe WL, Sharma S, Wu G, Yan C, Zhang H, Chung YH, Kim JA, Park NH, Song IH, Buetow KH (2010) Genetic variations at loci involved in the immune response are risk factors for hepatocellular carcinoma. Hepatology 52(6):2034–2043. doi: 10.1002/hep.23943PubMedCrossRefGoogle Scholar
  42. 42.
    Zhang H, Zhai Y, Hu Z, Wu C, Qian J, Jia W, Ma F, Huang W, Yu L, Yue W, Wang Z, Li P, Zhang Y, Liang R, Wei Z, Cui Y, Xie W, Cai M, Yu X, Yuan Y, Xia X, Zhang X, Yang H, Qiu W, Yang J, Gong F, Chen M, Shen H, Lin D, Zeng YX, He F, Zhou G (2010) Genome-wide association study identifies 1p36.22 as a new susceptibility locus for hepatocellular carcinoma in chronic hepatitis B virus carriers. Nat Genet 42(9):755–758. doi: ng.638[pii]10.1038/ng.638PubMedCrossRefGoogle Scholar
  43. 43.
    Kumar V, Kato N, Urabe Y, Takahashi A, Muroyama R, Hosono N, Otsuka M, Tateishi R, Omata M, Nakagawa H, Koike K, Kamatani N, Kubo M, Nakamura Y, Matsuda K (2011) Genome-wide association study identifies a susceptibility locus for HCV-induced hepatocellular carcinoma. Nat Genet 43(5):455–458. doi: ng.809[pii]10.1038/ng.809PubMedCrossRefGoogle Scholar
  44. 44.
    Miki D, Ochi H, Hayes CN, Abe H, Yoshima T, Aikata H, Ikeda K, Kumada H, Toyota J, Morizono T, Tsunoda T, Kubo M, Nakamura Y, Kamatani N, Chayama K (2011) Variation in the DEPDC5 locus is associated with progression to hepatocellular carcinoma in chronic hepatitis C virus carriers. Nat Genet 43(8):797–800. doi: ng.876[pii]10.1038/ng.876PubMedCrossRefGoogle Scholar
  45. 45.
    Villanueva A, Forns X, Llovet JM (2012) Molecular epidemiology in HCV-related hepatocellular carcinoma: first steps. J Hepatol. doi: S0168-8278(12)3-3[pii]10.1016/j.jhep.2012.01.002Google Scholar
  46. 46.
    Rodenhiser D, Mann M (2006) Epigenetics and human disease: translating basic biology into clinical applications. Can Med Assoc J 174(3):341–348. doi: 174/3/341[pii]10.1503/cmaj.050774CrossRefGoogle Scholar
  47. 47.
    Ducasse M, Brown MA (2006) Epigenetic aberrations and cancer. Mol Cancer 5:60. doi: 1476-4598-5-60[pii]10.1186/1476-4598-5-60PubMedCrossRefGoogle Scholar
  48. 48.
    Gibney ER, Nolan CM (2010) Epigenetics and gene expression. Heredity (Edinb) 105(1):4–13. doi: hdy201054[pii]10.1038/hdy.2010.54CrossRefGoogle Scholar
  49. 49.
    Sandoval J, Esteller M (2012) Cancer epigenomics: beyond genomics. Curr Opin Genet Dev 22(1):50–55. doi: S0959-437X(12)00019-6[pii]10.1016/j.gde.2012.02.008PubMedCrossRefGoogle Scholar
  50. 50.
    Sharma S, Kelly TK, Jones PA (2010) Epigenetics in cancer. Carcinogenesis 31(1):27–36. doi: bgp220[pii]10.1093/carcin/bgp220PubMedCrossRefGoogle Scholar
  51. 51.
    Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297. doi: S0092867404000455[pii]PubMedCrossRefGoogle Scholar
  52. 52.
    Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136(2):215–233. doi: S0092-8674(09)00008-7[pii]10.1016/j.cell.2009.01.002PubMedCrossRefGoogle Scholar
  53. 53.
    Liu X, Fortin K, Mourelatos Z (2008) MicroRNAs: biogenesis and molecular functions. Brain Pathol 18(1):113–121. doi: BPA121[pii]10.1111/j.1750-3639.2007.00121.xPubMedCrossRefGoogle Scholar
  54. 54.
    Ambros V (2004) The functions of animal microRNAs. Nature 431(7006):350–355. doi: nature02871[pii]10.1038/nature02871PubMedCrossRefGoogle Scholar
  55. 55.
    He L, Hannon GJ (2004) MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 5(7):522–531. doi: nrg1379[pii]10.1038/nrg1379PubMedCrossRefGoogle Scholar
  56. 56.
    Calin GA, Croce CM (2006) MicroRNA signatures in human cancers. Nat Rev Cancer 6(11):857–866. doi: nrc1997[pii]10.1038/nrc1997PubMedCrossRefGoogle Scholar
  57. 57.
    Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, Sweet-Cordero A, Ebert BL, Mak RH, Ferrando AA, Downing JR, Jacks T, Horvitz HR, Golub TR (2005) MicroRNA expression profiles classify human cancers. Nature 435(7043):834–838. doi: nature03702[pii]10.1038/nature03702PubMedCrossRefGoogle Scholar
  58. 58.
    Murakami Y, Yasuda T, Saigo K, Urashima T, Toyoda H, Okanoue T, Shimotohno K (2006) Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues. Oncogene 25(17):2537–2545. doi: 1209283[pii]10.1038/sj.onc.1209283PubMedCrossRefGoogle Scholar
  59. 59.
    Braconi C, Patel T (2008) MicroRNA expression profiling: a molecular tool for defining the phenotype of hepatocellular tumors. Hepatology 47(6):1807–1809. doi: 10.1002/hep.22326PubMedCrossRefGoogle Scholar
  60. 60.
    Budhu A, Jia HL, Forgues M, Liu CG, Goldstein D, Lam A, Zanetti KA, Ye QH, Qin LX, Croce CM, Tang ZY, Wang XW (2008) Identification of metastasis-related microRNAs in hepatocellular carcinoma. Hepatology 47(3):897–907. doi: 10.1002/hep.22160PubMedCrossRefGoogle Scholar
  61. 61.
    Ladeiro Y, Couchy G, Balabaud C, Bioulac-Sage P, Pelletier L, Rebouissou S, Zucman-Rossi J (2008) MicroRNA profiling in hepatocellular tumors is associated with clinical features and oncogene/tumor suppressor gene mutations. Hepatology 47(6):1955–1963. doi: 10.1002/hep.22256PubMedCrossRefGoogle Scholar
  62. 62.
    Li W, Xie L, He X, Li J, Tu K, Wei L, Wu J, Guo Y, Ma X, Zhang P, Pan Z, Hu X, Zhao Y, Xie H, Jiang G, Chen T, Wang J, Zheng S, Cheng J, Wan D, Yang S, Li Y, Gu J (2008) Diagnostic and prognostic implications of microRNAs in human hepatocellular carcinoma. Int J Cancer 123(7):1616–1622. doi: 10.1002/ijc.23693PubMedCrossRefGoogle Scholar
  63. 63.
    Wong QW, Lung RW, Law PT, Lai PB, Chan KY, To KF, Wong N (2008) MicroRNA-223 is commonly repressed in hepatocellular carcinoma and potentiates expression of Stathmin1. Gastroenterology 135(1):257–269. doi: S0016-5085(08)00628-8[pii]10.1053/j.gastro.2008.04.003PubMedCrossRefGoogle Scholar
  64. 64.
    Jiang J, Gusev Y, Aderca I, Mettler TA, Nagorney DM, Brackett DJ, Roberts LR, Schmittgen TD (2008) Association of microRNA expression in hepatocellular carcinomas with hepatitis infection, cirrhosis, and patient survival. Clin Cancer Res 14(2):419–427. doi: 14/2/419[pii]10.1158/1078-0432.CCR-07-0523PubMedCrossRefGoogle Scholar
  65. 65.
    Wang Y, Lee AT, Ma JZ, Wang J, Ren J, Yang Y, Tantoso E, Li KB, Ooi LL, Tan P, Lee CG (2008) Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target. J Biol Chem 283(19):13205–13215. doi: M707629200[pii]10.1074/jbc.M707629200PubMedCrossRefGoogle Scholar
  66. 66.
    Esquela-Kerscher A, Slack FJ (2006) Oncomirs – microRNAs with a role in cancer. Nat Rev Cancer 6(4):259–269. doi: nrc1840[pii]10.1038/nrc1840PubMedCrossRefGoogle Scholar
  67. 67.
    Ventura A, Jacks T (2009) MicroRNAs and cancer: short RNAs go a long way. Cell 136(4):586–591. doi: S0092-8674(09)00141-X[pii]10.1016/j.cell.2009.02.005PubMedCrossRefGoogle Scholar
  68. 68.
    Calin GA, Dumitru CD, Shimizu M, Bichi R, Zupo S, Noch E, Aldler H, Rattan S, Keating M, Rai K, Rassenti L, Kipps T, Negrini M, Bullrich F, Croce CM (2002) Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc Natl Acad Sci USA 99(24):15524–15529. doi: 242606799[pii]10.1073/pnas.242606799PubMedCrossRefGoogle Scholar
  69. 69.
    Calin GA, Sevignani C, Dumitru CD, Hyslop T, Noch E, Yendamuri S, Shimizu M, Rattan S, Bullrich F, Negrini M, Croce CM (2004) Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA 101(9):2999–3004. doi: 0307323101[pii]10.1073/pnas.0307323101PubMedCrossRefGoogle Scholar
  70. 70.
    Mott JL (2009) MicroRNAs involved in tumor suppressor and oncogene pathways: implications for hepatobiliary neoplasia. Hepatology 50(2):630–637. doi: 10.1002/hep.23010PubMedCrossRefGoogle Scholar
  71. 71.
    Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST, Patel T (2007) MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology 133(2):647–658. doi: S0016-5085(07)01002-5[pii]10.1053/j.gastro.2007.05.022PubMedCrossRefGoogle Scholar
  72. 72.
    Gramantieri L, Fornari F, Ferracin M, Veronese A, Sabbioni S, Calin GA, Grazi GL, Croce CM, Bolondi L, Negrini M (2009) MicroRNA-221 targets Bmf in hepatocellular carcinoma and correlates with tumor multifocality. Clin Cancer Res 15(16):5073–5081. doi: 1078-0432.CCR-09-0092[pii]10.1158/1078-0432.CCR-09-0092PubMedCrossRefGoogle Scholar
  73. 73.
    Fornari F, Gramantieri L, Ferracin M, Veronese A, Sabbioni S, Calin GA, Grazi GL, Giovannini C, Croce CM, Bolondi L, Negrini M (2008) MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma. Oncogene 27(43):5651–5661. doi: onc2008178[pii]10.1038/onc.2008.178PubMedCrossRefGoogle Scholar
  74. 74.
    Tannapfel A, Grund D, Katalinic A, Uhlmann D, Köckerling F, Haugwitz U, Wasner M, Hauss J, Engeland K, Wittekind C (2000) Decreased expression of p27 protein is associated with advanced tumor stage in hepatocellular carcinoma. Int J Cancer 89(4):350–355. doi:10.1002/1097-0215(20000720)89:4<350::AID-IJC6>3.0.CO;2-3 [pii]PubMedCrossRefGoogle Scholar
  75. 75.
    Ito Y, Takeda T, Sakon M, Tsujimoto M, Monden M, Matsuura N (2001) Expression of p57/Kip2 protein in hepatocellular carcinoma. Oncology 61(3):221–225. doi: 55378[pii]PubMedCrossRefGoogle Scholar
  76. 76.
    Nakai S, Masaki T, Shiratori Y, Ohgi T, Morishita A, Kurokohchi K, Watanabe S, Kuriyama S (2002) Expression of p57(KIP2) in hepatocellular carcinoma: relationship between tumor differentiation and patient survival. Int J Oncol 20(4):769–775PubMedGoogle Scholar
  77. 77.
    Liu WH, Yeh SH, Lu CC, Yu SL, Chen HY, Lin CY, Chen DS, Chen PJ (2009) MicroRNA-18a prevents estrogen receptor-alpha expression, promoting proliferation of hepatocellular carcinoma cells. Gastroenterology 136(2):683–693. doi: S0016-5085(08)01864-7[pii]10.1053/j.gastro.2008.10.029PubMedCrossRefGoogle Scholar
  78. 78.
    Gramantieri L, Ferracin M, Fornari F, Veronese A, Sabbioni S, Liu CG, Calin GA, Giovannini C, Ferrazzi E, Grazi GL, Croce CM, Bolondi L, Negrini M (2007) Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma. Cancer Res 67(13):6092–6099. doi: 67/13/6092[pii]10.1158/0008-5472.CAN-06-4607PubMedCrossRefGoogle Scholar
  79. 79.
    Lin CJ, Gong HY, Tseng HC, Wang WL, Wu JL (2008) miR-122 targets an anti-apoptotic gene, Bcl-w, in human hepatocellular carcinoma cell lines. Biochem Biophys Res Commun 375(3):315–320. doi: S-291X(08)01485-X[pii]10.1016/j.bbrc.2008.07.154PubMedCrossRefGoogle Scholar
  80. 80.
    Bai S, Nasser MW, Wang B, Hsu SH, Datta J, Kutay H, Yadav A, Nuovo G, Kumar P, Ghoshal K (2009) MicroRNA-122 inhibits tumorigenic properties of hepatocellular carcinoma cells and sensitizes these cells to sorafenib. J Biol Chem 284(46):32015–32027. doi: M109.016774[pii]10.1074/jbc.M109.016774PubMedCrossRefGoogle Scholar
  81. 81.
    Tsai WC, Hsu PW, Lai TC, Chau GY, Lin CW, Chen CM, Lin CD, Liao YL, Wang JL, Chau YP, Hsu MT, Hsiao M, Huang HD, Tsou AP (2009) MicroRNA-122, a tumor suppressor microRNA that regulates intrahepatic metastasis of hepatocellular carcinoma. Hepatology 49(5):1571–1582. doi: 10.1002/hep.22806PubMedCrossRefGoogle Scholar
  82. 82.
    Ji J, Shi J, Budhu A, Yu Z, Forgues M, Roessler S, Ambs S, Chen Y, Meltzer PS, Croce CM, Qin LX, Man K, Lo CM, Lee J, Ng IO, Fan J, Tang ZY, Sun HC, Wang XW (2009) MicroRNA expression, survival, and response to interferon in liver cancer. N Engl J Med 361(15):1437–1447. doi: 361/15/1437[pii]10.1056/NEJMoa0901282PubMedCrossRefGoogle Scholar
  83. 83.
    Mazzaferro V, Romito R, Schiavo M, Mariani L, Camerini T, Bhoori S, Capussotti L, Calise F, Pellicci R, Belli G, Tagger A, Colombo M, Bonino F, Majno P, Llovet JM, Force HIT (2006) Prevention of hepatocellular carcinoma recurrence with alpha-interferon after liver resection in HCV cirrhosis. Hepatology 44(6):1543–1554. doi: 10.1002/hep.21415PubMedCrossRefGoogle Scholar
  84. 84.
    Li S, Fu H, Wang Y, Tie Y, Xing R, Zhu J, Sun Z, Wei L, Zheng X (2009) MicroRNA-101 regulates expression of the v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) oncogene in human hepatocellular carcinoma. Hepatology 49(4):1194–1202. doi: 10.1002/hep.22757PubMedCrossRefGoogle Scholar
  85. 85.
    Su H, Yang JR, Xu T, Huang J, Xu L, Yuan Y, Zhuang SM (2009) MicroRNA-101, down-regulated in hepatocellular carcinoma, promotes apoptosis and suppresses tumorigenicity. Cancer Res 69(3):1135–1142. doi: 0008-5472.CAN-08-2886[pii]10.1158/0008-5472.CAN-08-2886PubMedCrossRefGoogle Scholar
  86. 86.
    Li D, Liu X, Lin L, Hou J, Li N, Wang C, Wang P, Zhang Q, Zhang P, Zhou W, Wang Z, Ding G, Zhuang SM, Zheng L, Tao W, Cao X (2011) MicroRNA-99a inhibits hepatocellular carcinoma growth and correlates with prognosis of patients with hepatocellular carcinoma. J Biol Chem 286(42):36677–36685. doi: M111.270561[pii]10.1074/jbc.M111.270561PubMedCrossRefGoogle Scholar
  87. 87.
    Chung GE, Yoon JH, Myung SJ, Lee JH, Lee SH, Lee SM, Kim SJ, Hwang SY, Lee HS, Kim CY (2010) High expression of microRNA-15b predicts a low risk of tumor recurrence following curative resection of hepatocellular carcinoma. Oncol Rep 23(1):113–119PubMedGoogle Scholar
  88. 88.
    Huang YS, Dai Y, Yu XF, Bao SY, Yin YB, Tang M, Hu CX (2008) Microarray analysis of microRNA expression in hepatocellular carcinoma and non-tumorous tissues without viral hepatitis. J Gastroenterol Hepatol 23(1):87–94. doi: JGH5223[pii]10.1111/j.1440-1746.2007.05223.xPubMedCrossRefGoogle Scholar
  89. 89.
    De Zhu J (2005) The altered DNA methylation pattern and its implications in liver cancer. Cell Res 15(4):272–280. doi: 10.1038/sj.cr.7290296PubMedCrossRefGoogle Scholar
  90. 90.
    Zhu J (2006) DNA methylation and hepatocellular carcinoma. J Hepatobiliary Pancreat Surg 13(4):265–273. doi: 10.1007/s00534-005-1054-4PubMedCrossRefGoogle Scholar
  91. 91.
    Tischoff I, Tannapfe A (2008) DNA methylation in hepatocellular carcinoma. World J Gastroenterol 14(11):1741–1748PubMedCrossRefGoogle Scholar
  92. 92.
    Kanai Y, Hirohashi S (2007) Alterations of DNA methylation associated with abnormalities of DNA methyltransferases in human cancers during transition from a precancerous to a malignant state. Carcinogenesis 28(12):2434–2442. doi: bgm206[pii]10.1093/carcin/bgm206PubMedCrossRefGoogle Scholar
  93. 93.
    Arai E, Ushijima S, Gotoh M, Ojima H, Kosuge T, Hosoda F, Shibata T, Kondo T, Yokoi S, Imoto I, Inazawa J, Hirohashi S, Kanai Y (2009) Genome-wide DNA methylation profiles in liver tissue at the precancerous stage and in hepatocellular carcinoma. Int J Cancer 125(12):2854–2862. doi: 10.1002/ijc.24708PubMedCrossRefGoogle Scholar
  94. 94.
    Lee HS, Kim BH, Cho NY, Yoo EJ, Choi M, Shin SH, Jang JJ, Suh KS, Kim YS, Kang GH (2009) Prognostic implications of and relationship between CpG island hypermethylation and repetitive DNA hypomethylation in hepatocellular carcinoma. Clin Cancer Res 15(3):812–820. doi: 15/3/812[pii]10.1158/1078-0432.CCR-08-0266PubMedCrossRefGoogle Scholar
  95. 95.
    Lin CH, Hsieh SY, Sheen IS, Lee WC, Chen TC, Shyu WC, Liaw YF (2001) Genome-wide hypomethylation in hepatocellular carcinogenesis. Cancer Res 61(10):4238–4243PubMedGoogle Scholar
  96. 96.
    Gao W, Kondo Y, Shen L, Shimizu Y, Sano T, Yamao K, Natsume A, Goto Y, Ito M, Murakami H, Osada H, Zhang J, Issa JP, Sekido Y (2008) Variable DNA methylation patterns associated with progression of disease in hepatocellular carcinomas. Carcinogenesis 29(10):1901–1910. doi: bgn170[pii]10.1093/carcin/bgn170PubMedCrossRefGoogle Scholar
  97. 97.
    Nishida N, Nagasaka T, Nishimura T, Ikai I, Boland CR, Goel A (2008) Aberrant methylation of multiple tumor suppressor genes in aging liver, chronic hepatitis, and hepatocellular carcinoma. Hepatology 47(3):908–918. doi: 10.1002/hep.22110PubMedCrossRefGoogle Scholar
  98. 98.
    Rivenbark AG, Coleman WB (2007) The use of epigenetic biomarkers for preclinical detection of hepatocellular carcinoma: potential for noninvasive screening of high-risk populations. Clin Cancer Res 13(8):2309–2312. doi: 13/8/2309[pii]10.1158/1078-0432.CCR-07-0086PubMedCrossRefGoogle Scholar
  99. 99.
    Yamada S, Nomoto S, Fujii T, Takeda S, Kanazumi N, Sugimoto H, Nakao A (2007) Frequent promoter methylation of M-cadherin in hepatocellular carcinoma is associated with poor prognosis. Anticancer Res 27(4B):2269–2274PubMedGoogle Scholar
  100. 100.
    Yan Q, Zhang ZF, Chen XP, Gutmann DH, Xiong M, Xiao ZY, Huang ZY (2008) Reduced T-cadherin expression and promoter methylation are associated with the development and progression of hepatocellular carcinoma. Int J Oncol 32(5):1057–1063PubMedGoogle Scholar
  101. 101.
    Jung JK, Arora P, Pagano JS, Jang KL (2007) Expression of DNA methyltransferase 1 is activated by hepatitis B virus X protein via a regulatory circuit involving the p16INK4a-cyclin D1-CDK 4/6-pRb-E2F1 pathway. Cancer Res 67(12):5771–5778. doi: 67/12/5771[pii]10.1158/0008-5472.CAN-07-0529PubMedCrossRefGoogle Scholar
  102. 102.
    Yang B, Guo M, Herman JG, Clark DP (2003) Aberrant promoter methylation profiles of tumor suppressor genes in hepatocellular carcinoma. Am J Pathol 163(3):1101–1107. doi: S0002-9440(10)63469-4[pii]10.1016/S0002-9440(10)63469-4PubMedCrossRefGoogle Scholar
  103. 103.
    Calvisi DF, Ladu S, Gorden A, Farina M, Lee JS, Conner EA, Schroeder I, Factor VM, Thorgeirsson SS (2007) Mechanistic and prognostic significance of aberrant methylation in the molecular pathogenesis of human hepatocellular carcinoma. J Clin Invest 117(9):2713–2722. doi: 10.1172/JCI31457PubMedCrossRefGoogle Scholar
  104. 104.
    Saito Y, Kanai Y, Nakagawa T, Sakamoto M, Saito H, Ishii H, Hirohashi S (2003) Increased protein expression of DNA methyltransferase (DNMT) 1 is significantly correlated with the malignant potential and poor prognosis of human hepatocellular carcinomas. Int J Cancer 105(4):527–532. doi: 10.1002/ijc.11127PubMedCrossRefGoogle Scholar
  105. 105.
    Oh BK, Kim H, Park HJ, Shim YH, Choi J, Park C, Park YN (2007) DNA methyltransferase expression and DNA methylation in human hepatocellular carcinoma and their clinicopathological correlation. Int J Mol Med 20(1):65–73PubMedGoogle Scholar
  106. 106.
    Cedar H, Bergman Y (2009) Linking DNA methylation and histone modification: patterns and paradigms. Nat Rev Genet 10(5):295–304. doi: nrg2540[pii]10.1038/nrg2540PubMedCrossRefGoogle Scholar
  107. 107.
    Zhang K, Dent SY (2005) Histone modifying enzymes and cancer: going beyond histones. J Cell Biochem 96(6):1137–1148. doi: 10.1002/jcb.20615PubMedCrossRefGoogle Scholar
  108. 108.
    Santos-Rosa H, Caldas C (2005) Chromatin modifier enzymes, the histone code and cancer. Eur J Cancer 41(16):2381–2402. doi: S0959-8049(05)00710-0[pii]10.1016/j.ejca.2005.08.010PubMedCrossRefGoogle Scholar
  109. 109.
    Esteller M (2008) Epigenetics in cancer. N Engl J Med 358(11):1148–1159. doi: 358/11/1148[pii]10.1056/NEJMra072067PubMedCrossRefGoogle Scholar
  110. 110.
    Bernstein BE, Meissner A, Lander ES (2007) The mammalian epigenome. Cell 128(4):669–681. doi: S0092-8674(07)00128-6[pii]10.1016/j.cell.2007.01.033PubMedCrossRefGoogle Scholar
  111. 111.
    Kondo Y, Shen L, Suzuki S, Kurokawa T, Masuko K, Tanaka Y, Kato H, Mizuno Y, Yokoe M, Sugauchi F, Hirashima N, Orito E, Osada H, Ueda R, Guo Y, Chen X, Issa JP, Sekido Y (2007) Alterations of DNA methylation and histone modifications contribute to gene silencing in hepatocellular carcinomas. Hepatol Res 37(11):974–983. doi: HEP141[pii]10.1111/j.1872-034X.2007.00141.xPubMedCrossRefGoogle Scholar
  112. 112.
    Pogribny IP, Ross SA, Tryndyak VP, Pogribna M, Poirier LA, Karpinets TV (2006) Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4-20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats. Carcinogenesis 27(6):1180–1186. doi: bgi364[pii]10.1093/carcin/bgi364PubMedCrossRefGoogle Scholar
  113. 113.
    Toh Y, Nicolson GL (2009) The role of the MTA family and their encoded proteins in human cancers: molecular functions and clinical implications. Clin Exp Metastasis 26(3):215–227. doi: 10.1007/s10585-008-9233-8PubMedCrossRefGoogle Scholar
  114. 114.
    Lee H, Ryu SH, Hong SS, Seo DD, Min HJ, Jang MK, Kwon HJ, Yu E, Chung YH, Kim KW (2009) Overexpression of metastasis-associated protein 2 is associated with hepatocellular carcinoma size and differentiation. J Gastroenterol Hepatol 24(8):1445–1450. doi: JGH5965[pii]10.1111/j.1440-1746.2009.05965.xPubMedCrossRefGoogle Scholar
  115. 115.
    Hamatsu T, Rikimaru T, Yamashita Y, Aishima S, Tanaka S, Shirabe K, Shimada M, Toh Y, Sugimachi K (2003) The role of MTA1 gene expression in human hepatocellular carcinoma. Oncol Rep 10(3):599–604PubMedGoogle Scholar
  116. 116.
    Moon WS, Chang K, Tarnawski AS (2004) Overexpression of metastatic tumor antigen 1 in hepatocellular carcinoma: relationship to vascular invasion and estrogen receptor-alpha. Hum Pathol 35(4):424–429. doi: S0046817703006713[pii]PubMedCrossRefGoogle Scholar
  117. 117.
    Yoo YG, Na TY, Seo HW, Seong JK, Park CK, Shin YK, Lee MO (2008) Hepatitis B virus X protein induces the expression of MTA1 and HDAC1, which enhances hypoxia signaling in hepatocellular carcinoma cells. Oncogene 27(24):3405–3413. doi: 1211000[pii]10.1038/sj.onc.1211000PubMedCrossRefGoogle Scholar
  118. 118.
    Ryu SH, Chung YH, Lee H, Kim JA, Shin HD, Min HJ, Seo DD, Jang MK, Yu E, Kim KW (2008) Metastatic tumor antigen 1 is closely associated with frequent postoperative recurrence and poor survival in patients with hepatocellular carcinoma. Hepatology 47(3):929–936. doi: 10.1002/hep.22124PubMedCrossRefGoogle Scholar
  119. 119.
    Rogers S, Girolami M, Kolch W, Waters KM, Liu T, Thrall B, Wiley HS (2008) Investigating the correspondence between transcriptomic and proteomic expression profiles using coupled cluster models. Bioinformatics 24(24):2894–2900. doi: btn553[pii]10.1093/bioinformatics/btn553PubMedCrossRefGoogle Scholar
  120. 120.
    Kim W, Oe Lim S, Kim JS, Ryu YH, Byeon JY, Kim HJ, Kim YI, Heo JS, Park YM, Jung G (2003) Comparison of proteome between hepatitis B virus- and hepatitis C virus-associated hepatocellular carcinoma. Clin Cancer Res 9(15):5493–5500PubMedGoogle Scholar
  121. 121.
    Li C, Tan YX, Zhou H, Ding SJ, Li SJ, Ma DJ, Man XB, Hong Y, Zhang L, Li L, Xia QC, Wu JR, Wang HY, Zeng R (2005) Proteomic analysis of hepatitis B virus-associated hepatocellular carcinoma: identification of potential tumor markers. Proteomics 5(4):1125–1139. doi: 10.1002/pmic.200401141PubMedCrossRefGoogle Scholar
  122. 122.
    Li C, Ruan HQ, Liu YS, Xu MJ, Dai J, Sheng QH, Tan YX, Yao ZZ, Wang HY, Wu JR, Zeng R (2012) Quantitative proteomics reveal up-regulated protein expression of the SET complex associated with hepatocellular carcinoma. J Proteome Res 11(2):871–885. doi: 10.1021/pr2006999PubMedCrossRefGoogle Scholar
  123. 123.
    Roessler S, Long EL, Budhu A, Chen Y, Zhao X, Ji J, Walker R, Jia HL, Ye QH, Qin LX, Tang ZY, He P, Hunter KW, Thorgeirsson SS, Meltzer PS, Wang XW (2011) Integrative genomic identification of genes on 8p associated with hepatocellular carcinoma progression and patient survival. Gastroenterology. doi: S0016-5085(11)01771-9[pii]10.1053/j.gastro.2011.12.039Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Rosina Maria Critelli
    • 1
  • Elisabetta Cariani
    • 2
  • Erica Villa
    • 1
    • 3
  1. 1.Division of Gastroenterology, Department of Internal MedicineUniversity of Modena and Reggio EmiliaModenaItaly
  2. 2.Clinical PathologyOspedale Civile S. Agostino-EstenseModenaItaly
  3. 3.Department of GastroenterologyAzienda Ospedaliero-Universitaria di Modena & University of Modena and Reggio EmiliaModenaItaly

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