Crosstalk of Ras and Rho: activation of RhoA abates Kras-induced liver tumorigenesis in transgenic zebrafish models

Chew, T.W., Liu, X.J., Liu, L., Spitsbergen, J.M., Gong, Z., and Low, B.C.
Oncogene   33(21): 2717-27 (Journal)
Registered Authors
Gong, Zhiyuan, Liu, Lihui, Liu, Xingjun, Low, Boon Chaun, Spitsbergen, Jan
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Carcinogenesis/metabolism*
  • Carcinoma, Hepatocellular/enzymology*
  • Carcinoma, Hepatocellular/genetics
  • Cell Proliferation
  • Enzyme Activation
  • Hepatocytes/enzymology
  • Humans
  • Liver/enzymology
  • Liver/pathology
  • Liver Neoplasms, Experimental/enzymology*
  • Liver Neoplasms, Experimental/genetics
  • MAP Kinase Signaling System
  • Monomeric GTP-Binding Proteins/metabolism*
  • Mutation, Missense
  • Proto-Oncogene Proteins c-akt/metabolism
  • Proto-Oncogene Proteins p21(ras)/genetics*
  • Zebrafish
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism*
23812423 Full text @ Oncogene

RAS and Rho small GTPases are key molecular switches that control cell dynamics, cell growth and tissue development through their distinct signaling pathways. Although much has been learnt about their individual functions in both cell and animal models, the physiological and pathophysiological consequences of their signaling crosstalk in multi-cellular context in vivo remain largely unknown, especially in liver development and liver tumorigenesis. Furthermore, the roles of RhoA in RAS-mediated transformation and their crosstalk in vitro remain highly controversial. When challenged with carcinogens, zebrafish developed liver cancer that resembles the human liver cancer both molecularly and histopathologically. Capitalizing on the growing importance and relevance of zebrafish (Danio rerio) as an alternate cancer model, we have generated liver-specific, Tet-on-inducible transgenic lines expressing oncogenic KrasG12V, RhoA, constitutively active RhoAG14V or dominant-negative RhoAT19N. Double-transgenic lines expressing KrasG12V with one of the three RhoA genes were also generated. Based on quantitative bioimaging and molecular markers for genetic and signaling aberrations, we showed that the induced expression of oncogenic Kras during early development led to liver enlargement and hepatocyte proliferation, associated with elevated Erk phosphorylation, activation of Akt2 and modulation of its two downstream targets, p21Cip and S6 kinase. Such an increase in liver size and Akt2 expression was augmented by dominant-negative RhoAT19N, but was abrogated by the constitutive-active RhoAG14V. Consequently, induced expression of the oncogenic Kras in adult transgenic fish led to the development of hepatocellular carcinomas. Survival studies further revealed that the co-expression of dominant-negative RhoAT19N with oncogenic Kras increased the mortality rate compared with the other single or double-transgenic lines. This study provides evidence of the previously unappreciated signaling crosstalk between Kras and RhoA in regulating liver overgrowth and liver tumorigenesis. Our results also implicate that activating Rho could be beneficial to suppress the Kras-induced liver malignancies.

Genes / Markers
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Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes