A High Level of Liver-specific Expression of Oncogenic Kras(V12) Drives Robust Liver Tumorigenesis in Transgenic Zebrafish

Overview

Journal Dis Model Mech

Specialty General Medicine

Date 2011 Jul 7

PMID 21729876

Citations 56

Authors

Anh Tuan Nguyen

Alexander Emelyanov

Chor Hui Vivien Koh

Jan M Spitsbergen

Siew Hong Lam

Sinnakaruppan Mathavan

Serguei Parinov

Zhiyuan Gong

Affiliations

Soon will be listed here.

Abstract

Human liver cancer is one of the deadliest cancers worldwide, with hepatocellular carcinoma (HCC) being the most common type. Aberrant Ras signaling has been implicated in the development and progression of human HCC, but a complete understanding of the molecular mechanisms of this protein in hepatocarcinogenesis remains elusive. In this study, a stable in vivo liver cancer model using transgenic zebrafish was generated to elucidate Ras-driven tumorigenesis in HCC. Using the liver-specific fabp10 (fatty acid binding protein 10) promoter, we overexpressed oncogenic kras(V12) specifically in the transgenic zebrafish liver. Only a high level of kras(V12) expression initiated liver tumorigenesis, which progressed from hyperplasia to benign and malignant tumors with activation of the Ras-Raf-MEK-ERK and Wnt-β-catenin pathways. Histological diagnosis of zebrafish tumors identified HCC as the main lesion. The tumors were invasive and transplantable, indicating malignancy of these HCC cells. Oncogenic kras(V12) was also found to trigger p53-dependent senescence as a tumor suppressive barrier in the pre-neoplastic stage. Microarray analysis of zebrafish liver hyperplasia and HCC uncovered the deregulation of several stage-specific and common biological processes and signaling pathways responsible for kras(V12)-driven liver tumorigenesis that recapitulated the molecular hallmarks of human liver cancer. Cross-species comparisons of cancer transcriptomes further defined a HCC-specific gene signature as well as a liver cancer progression gene signature that are evolutionarily conserved between human and zebrafish. Collectively, our study presents a comprehensive portrait of molecular mechanisms during progressive Ras-induced HCC. These observations indicate the validity of our transgenic zebrafish to model human liver cancer, and this model might act as a useful platform for drug screening and identifying new therapeutic targets.

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References

Torimura T, Ueno T, Kin M, Harada R, Taniguchi E, Nakamura T . Overexpression of angiopoietin-1 and angiopoietin-2 in hepatocellular carcinoma. J Hepatol. 2004; 40(5):799-807. DOI: 10.1016/j.jhep.2004.01.027. View

Markiewski M, DeAngelis R, Benencia F, Ricklin-Lichtsteiner S, Koutoulaki A, Gerard C . Modulation of the antitumor immune response by complement. Nat Immunol. 2008; 9(11):1225-35. PMC: 2678913. DOI: 10.1038/ni.1655. View

Hayashi J, Aoki H, Kajino K, Moriyama M, Arakawa Y, Hino O . Hepatitis C virus core protein activates the MAPK/ERK cascade synergistically with tumor promoter TPA, but not with epidermal growth factor or transforming growth factor alpha. Hepatology. 2000; 32(5):958-61. DOI: 10.1053/jhep.2000.19343. View

Wei Y, Tran Van Nhieu J, Prigent S, Srivatanakul P, Tiollais P, Buendia M . Altered expression of E-cadherin in hepatocellular carcinoma: correlations with genetic alterations, beta-catenin expression, and clinical features. Hepatology. 2002; 36(3):692-701. DOI: 10.1053/jhep.2002.35342. View

Amatruda J, Patton E . Genetic models of cancer in zebrafish. Int Rev Cell Mol Biol. 2008; 271:1-34. DOI: 10.1016/S1937-6448(08)01201-X. View

Xue W, Zender L, Miething C, Dickins R, Hernando E, Krizhanovsky V . Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature. 2007; 445(7128):656-60. PMC: 4601097. DOI: 10.1038/nature05529. View

Lam S, Gong Z . Modeling liver cancer using zebrafish: a comparative oncogenomics approach. Cell Cycle. 2006; 5(6):573-7. DOI: 10.4161/cc.5.6.2550. View

Schmidt C, Mckillop I, Cahill P, Sitzmann J . Increased MAPK expression and activity in primary human hepatocellular carcinoma. Biochem Biophys Res Commun. 1997; 236(1):54-8. DOI: 10.1006/bbrc.1997.6840. View

Parinov S, Kondrichin I, Korzh V, Emelyanov A . Tol2 transposon-mediated enhancer trap to identify developmentally regulated zebrafish genes in vivo. Dev Dyn. 2004; 231(2):449-59. DOI: 10.1002/dvdy.20157. View

10.

Donehower L, Harvey M, Slagle B, McArthur M, Montgomery Jr C, Butel J . Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature. 1992; 356(6366):215-21. DOI: 10.1038/356215a0. View

11.

Korzh S, Pan X, Garcia-Lecea M, Winata C, Pan X, Wohland T . Requirement of vasculogenesis and blood circulation in late stages of liver growth in zebrafish. BMC Dev Biol. 2008; 8:84. PMC: 2564926. DOI: 10.1186/1471-213X-8-84. View

12.

Di Micco R, Fumagalli M, Cicalese A, Piccinin S, Gasparini P, Luise C . Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature. 2006; 444(7119):638-42. DOI: 10.1038/nature05327. View

13.

Lee J, Chu I, Mikaelyan A, Calvisi D, Heo J, Reddy J . Application of comparative functional genomics to identify best-fit mouse models to study human cancer. Nat Genet. 2004; 36(12):1306-11. DOI: 10.1038/ng1481. View

14.

Mizgirev I, Revskoy S . Generation of clonal zebrafish lines and transplantable hepatic tumors. Nat Protoc. 2010; 5(3):383-94. DOI: 10.1038/nprot.2010.8. View

15.

Harada N, Oshima H, Katoh M, Tamai Y, Oshima M, Taketo M . Hepatocarcinogenesis in mice with beta-catenin and Ha-ras gene mutations. Cancer Res. 2004; 64(1):48-54. DOI: 10.1158/0008-5472.can-03-2123. View

16.

Calvisi D, Ladu S, Gorden A, Farina M, Conner E, Lee J . Ubiquitous activation of Ras and Jak/Stat pathways in human HCC. Gastroenterology. 2006; 130(4):1117-28. DOI: 10.1053/j.gastro.2006.01.006. View

17.

Wurmbach E, Chen Y, Khitrov G, Zhang W, Roayaie S, Schwartz M . Genome-wide molecular profiles of HCV-induced dysplasia and hepatocellular carcinoma. Hepatology. 2007; 45(4):938-47. DOI: 10.1002/hep.21622. View

18.

Lam S, Lian Wu Y, Vega V, Miller L, Spitsbergen J, Tong Y . Conservation of gene expression signatures between zebrafish and human liver tumors and tumor progression. Nat Biotechnol. 2005; 24(1):73-5. DOI: 10.1038/nbt1169. View

19.

Berghmans S, Murphey R, Wienholds E, Neuberg D, Kutok J, Fletcher C . tp53 mutant zebrafish develop malignant peripheral nerve sheath tumors. Proc Natl Acad Sci U S A. 2005; 102(2):407-12. PMC: 544293. DOI: 10.1073/pnas.0406252102. View

20.

Yokoyama Y, Kuramitsu Y, Takashima M, Iizuka N, Terai S, Oka M . Protein level of apolipoprotein E increased in human hepatocellular carcinoma. Int J Oncol. 2006; 28(3):625-31. View