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Conditional expression of mutated K-ras accelerates intestinal tumorigenesis in Msh2-deficient mice

Oncogene volume 26, pages 4415–4427 (2007)Cite this article

Abstract

K-ras mutation occurs in 40–50% of human colorectal adenomas and carcinomas, but its contribution to intestinal tumorigenesis in vivo is unclear. We developed K-rasV12 transgenic mice that were crossed with Ah-Cre mice to generate K-rasV12/Cre mice, which showed β-naphthoflavone-induction of Cre-mediated LoxP recombination that activated intestinal expression of K-rasV12 4A and 4B transcripts and proteins. Only very occasional intestinal adenomas were observed in β-naphthoflavone-treated K-rasV12/Cre mice aged up to 2 years, suggesting that mutated K-ras expression alone does not significantly initiate intestinal tumourigenesis. To investigate the effects of mutated K-ras on DNA mismatch repair (MMR)-deficient intestinal tumour formation, these mice were crossed with Msh2−/− mice to generate K-rasV12/Cre/Msh2−/− offspring. After β-naphthoflavone treatment, K-rasV12/Cre/Msh2−/− mice showed reduced average lifespan of 17.3±5.0 weeks from 26.9±6.8 (control Msh2−/− mice) (P<0.01). They demonstrated increased adenomas in the small intestine from 1.41 (Msh2−/− controls) to 7.75 per mouse (increased fivefold, P<0.01). In the large intestine, very few adenomas were found in Msh2−/− mice (0.13 per mouse) whereas K-rasV12/Cre/Msh2−/− mice produced 2.70 adenomas per mouse (increased 20-fold, P<0.01). Over 80% adenomas from K-rasV12/Cre/Msh2−/− mice showed transgene recombination with expression of K-rasV12 4A and 4B transcripts and proteins. Sequencing of endogenous murine K-ras showed mutations in two out of 10 tumours examined from Msh2−/− mice, but no mutations in 17 tumours from K-rasV12/Cre/Msh2−/− mice. Expression of K-rasV12 in tumours caused activation of the mitogen-activated protein kinase and Akt/protein kinase B signaling pathways, demonstrated by phosphorylation of p44MAPK, Akt and GSK3β, as well as transcriptional upregulation of Pem, Tcl-1 and Trap1a genes (known targets of K-rasV12 expression in stem cells). Thus, mutated K-ras cooperates synergistically with MMR deficiency to accelerate intestinal tumorigenesis, particularly in the large intestine.

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Acknowledgements

We thank Clive Lebozer and Xinping Gong (Department of Pathology, University of Cambridge) for technical assistance. This work was supported by grants from Cancer Research UK and the Wellcome Trust.

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Authors and Affiliations

  1. Department of Pathology, Addenbrooke's Hospital, University of Cambridge, Cambridge, UK

    F Luo, D G Brooks, H Ye, R Hamoudi, G Poulogiannis & M J Arends

  2. Sir Alastair Currie Cancer Research UK Laboratory, Molecular Medicine Centre, The University of Edinburgh, Western General Hospital, Edinburgh, UK

    C E Patek

  3. Department of Oncology, Cambridge Institute of Medical Research, Addenbrooke's Hospital Campus, University of Cambridge, Cambridge, UK

    D J Winton

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  1. F Luo
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Correspondence to M J Arends.

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Luo, F., Brooks, D., Ye, H. et al. Conditional expression of mutated K-ras accelerates intestinal tumorigenesis in Msh2-deficient mice. Oncogene 26, 4415–4427 (2007). https://doi.org/10.1038/sj.onc.1210231

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