Skip to main content
SpringerLink
Log in

A Mouse Model for Breast Cancer Induced by Amplification and Overexpression of the neu Promoter and Transgene

  • Original Articles
  • Published:
Molecular Medicine Aims and scope Submit manuscript

Abstract

Background

ErbB-2 is a critical oncogenic marker in human breast cancer. Its appearance correlates with poor prognosis and it is, therefore, an important target for physiologic investigation and therapeutic intervention. With this in mind, we have created and characterized two mouse breast cancer models that express rat wild type neu, the homologue of ErbB-2, and rat mutant neu under the control of the normal mouse neu promoter. These models in which the copy number of the neu gene is moderately amplified should more closely parallel the expression pattern of ErbB-2 seen in some cases of human breast cancer.

Materials and Methods

Transgenic mouse models were constructed by injecting one of the two pronuclei of a fertilized FVB/n egg and implanting it into a pseudopregnant Swiss/Webster mouse. Tissue expression was analyzed through the use of reverse transcription polymerase chain reaction and mammary histopathology examined by fixing, staining and mounting of the entire gland.

Results

In the former wild type model, we show that low level, long term expression of neu leads to abnormal lobuloalveolar development in virginal glands and incomplete regression in multiparous glands. Malignant foci form following multiple rounds of pregnancy and regression. In the latter model, a similarly directed transgene carrying the constitutively activated, mutant form of the rat neu gene, a stronger but similar phenotype is displayed.

Conclusion

Evidently minor perturbations in amplified neu expression are sufficient to alter mammary development and induce malignant transformation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Shih C, Padhy LC, Murray M, Weinberg RA. (1981) Transforming genes of carcinomas and neuroblastomas introduced into mouse fibroblasts. Nature 290: 261–264.

    Article  CAS  PubMed  Google Scholar 

  2. King CR, Kraus MH, Aaronson SA. (1985) Amplification of a novel v-erbB-related gene in a human mammary carcinoma. Science 229: 974–976.

    Article  CAS  PubMed  Google Scholar 

  3. Coussens L, Yang-Feng TL, Liao YC, et al. (1985) Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science 230: 1132–1139.

    Article  CAS  PubMed  Google Scholar 

  4. Semba K, Kamata N, Toyoshima K, Yamamoto T. (1985) A v-erbB-related protooncogene, c-erbB-2, is distinct from the c-erbB-1/epidermal growth factor-receptor gene and is amplified in a human salivary gland adenocarcinoma. Proc. Natl. Acad. Sci. U.S.A. 82: 6497–6501.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Zeillinger R, Kury F, Czerwenka K, et al. (1989) HER-2 amplification, steroid receptors and epidermal growth factor receptor in primary breast cancer. Oncogene 4: 109–114.

    PubMed  CAS  Google Scholar 

  6. Prigent SA, Lemoine NR. (1992) The type 1 (EGFR-related) family of growth factor receptors and their ligands. Prog. Growth Factor Res. 4: 1–24.

    Article  CAS  PubMed  Google Scholar 

  7. Slamon DJ, Godolphin W, Jones LA, et al. (1989) Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 244: 707–712.

    Article  CAS  PubMed  Google Scholar 

  8. Parkes HC, Lillycrop K, Howell A, Craig RK. (1990) C-erbB2 mRNA expression in human breast tumours: comparison with c-erbB2 DNA amplification and correlation with prognosis. Br. J. Cancer 61: 39–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Heintz NH, Leslie KO, Rogers LA, Howard PL. (1990) Amplification of the c-erb B-2 oncogene and prognosis of breast adenocarcinoma. Arch. Pathol. Lab. Med. 114: 160–163.

    PubMed  CAS  Google Scholar 

  10. Tiwari RK, Borgen PI, Wong GY, Cordon CC, Osborne MP. (1992) HER-2/neu amplification and overexpression in primary human breast cancer is associated with early metastasis. Anticancer Res. 12: 419–425.

    PubMed  CAS  Google Scholar 

  11. Makar A, Desmedt EJ, DePotter CT, Vanderheyden JD, Schatteman ES. (1990) Neu oncogene in breast cancer and its possible association with the risk of distant metastases. Acta Oncologica 29: 931–934.

    Article  CAS  PubMed  Google Scholar 

  12. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. (1987) Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235: 177–182.

    Article  CAS  PubMed  Google Scholar 

  13. Persons DL, Borelli KA, Hsu PH. (1997) Quantitation of HER-2/neu and c-myc gene amplification in breast carcinoma using fluorescence in situ hybridization. Mod. Pathol. 10: 720–727.

    PubMed  CAS  Google Scholar 

  14. Berger MS, Locher GW, Saurer S, et al. (1988) Correlation of c-erbB-2 gene amplification and protein expression in human breast carcinoma with nodal status and nuclear grading. Cancer Res. 48: 1238–1243.

    PubMed  CAS  Google Scholar 

  15. Hudziak RM, Schlessinger J, Ullrich A. (1987) Increased expression of the putative growth factor receptor p185HER2 causes transformation and tumorigenesis of NIH 3T3 cells. Proc. Natl. Acad. Sci. U.S.A. 84: 7159–7163.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Hung MC, Schechter AL, Chavray PY, Stern DF, Weinberg RA. (1986) Molecular cloning of the neu gene: absence of gross structural alteration in oncogenic alleles. Proc. Natl. Acad. Sci. U.S.A. 83: 261–264.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Di Marco E, Pierce JH, Knicley CL, Di Fiore PP. (1990) Transformation of NIH 3T3 cells by overexpression of the normal coding sequence of the rat neu gene. Mol. Cell Biol. 10: 3247–3252.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Muller WJ, Sinn E, Pattengale PK, Wallace R, Leder P. (1988) Single-step induction of mammary adenocarcinoma in transgenic mice bearing the activated c-neu oncogene. Cell 54: 105–115.

    Article  CAS  PubMed  Google Scholar 

  19. Guy CT, Webster MA, Schaller M, Parsons TJ, Cardiff RD, Muller WJ. (1992) Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease. Proc. Natl. Acad. Sci. U.S.A. 89: 10578–10582.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. White MR, Hung MC. (1992) Cloning and characterization of the mouse neu promoter. Oncogene 7: 677–683.

    PubMed  CAS  Google Scholar 

  21. Cardiff RD, Muller WJ. (1993) Transgenic mouse models of mammary tumorigenesis. Cancer Surv. 16: 97–113.

    PubMed  CAS  Google Scholar 

  22. Pittius CW, Sankaran L, Topper YJ, Henninghausen L. (1988) Comparison of the regulation of the whey acidic protein gene with that of a hybrid gene containing the whey acidic protein gene promoter in transgenic mice. Mol. Endocrinol. 2: 1027–1032.

    Article  CAS  PubMed  Google Scholar 

  23. Knepper JE, Medina D, Butel JS. (1986) Differential expression of endogenouse mouse mammary tumor virus genes during development of the BALB/c mammary gland. J. Virol. 59: 518–521.

    PubMed  PubMed Central  CAS  Google Scholar 

  24. Schroeder JA, Lee DC. (1998) Dynamic expression and activation of ERBB receptors in the developing mouse mammary gland. Cell Growth Differ. 9: 451–464.

    PubMed  CAS  Google Scholar 

  25. Flanagan JG, Leder P. (1988) neu Protooncogene fused to an immunoglobulin heavy chain gene requires immunoglobulin light chain for cell surface expression and oncogenic transformation. Proc. Natl. Acad. Sci. U.S.A. 85: 8057–8061.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Dougall WC, Qian X, Peterson NC, Miller MJ, Samanta A, Green MI. (1994) The neu oncogene: signal transduction pathways, transformation mechanisms and evolving therapies. Oncogene 9: 2109–2123.

    PubMed  CAS  Google Scholar 

  27. Sympson CJ, Talhouk RS, Alexander CM, et al. (1994) Targeted expression of stromelysin-1 in mammary gland provides evidence for a role of proteinases in branching morphogenesis and the requirement for an intact basement membrane for tissue-specific gene expression. J. Cell Biol. 125: 681–693.

    Article  CAS  PubMed  Google Scholar 

  28. Daniel CW, Silberstein GB. (1987) In: Neville MC, Daniels CW (eds.) The Mammary Gland Development, Regulation, and Function. Plenum Press, New York, N.Y., pp. 3–36.

  29. Borellini F, Oka T. (1989) Growth control and differentiation in mammary epithelial cells. Environ. Health Perspect. 80: 85–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Lee M, Kim H, Jeon D, et al. (1996) Iron metabolism-related genes and mitochondrial genes are induced during involution of mouse mammary gland. Biochem. Biophys. Res. Commun. 224: 164–168.

    Article  CAS  PubMed  Google Scholar 

  31. Teng C, Pentecost BT, Chen YH, Newbold RR, Eddy EM, McLachlan JA. (1989) Lactotransferrin gene expression in the mouse uterus and mammary gland. Endocrinology 124: 992–999.

    Article  CAS  PubMed  Google Scholar 

  32. Strange R, Li F, Saurer S, Burkhardt A, Fris RR. (1992) Apoptotic cell death and tissue remodelling during mouse mammary gland involution. Development 115: 49–58.

    PubMed  CAS  Google Scholar 

  33. Ross JS, Fletcher JA. (1998) The HER-2/neu oncogene in breast cancer: prognostic factor, predictive factor, and target for therapy. Stem Cells 16: 413–428.

    Article  CAS  PubMed  Google Scholar 

  34. Revillion F, Bonneterre J, Peyrat JP. (1998) ERBB2 oncogene in human breast cancer and its clinical significance. Eur. J. Cancer 34: 791–808.

    Article  CAS  PubMed  Google Scholar 

  35. Kokai Y, Cohen JA, Drebin JA, Greene MI. (1987) Stage- and tissue-specific expression of the neu oncogene in rat development. Proc. Natl. Acad. Sci. U.S.A. 84: 8498–8501.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Press MF, Cordon CC, Slamon DJ. (1990) Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues. Oncogene 5: 953–962.

    PubMed  CAS  Google Scholar 

  37. MacMahon B, Cole P, Lin TM, et al. (1970) Age at first birth and breast cancer risk. Bull. World Health Organ. 43: 209–221.

    PubMed  PubMed Central  CAS  Google Scholar 

  38. Siegel PM, Dankort DL, Hardy WR, Muller WJ. (1994) Novel activating mutations in the neu proto-oncogene involved in induction of mammary tumors. Mol. Cell Biol. 14: 7068–7077.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

We would like to thank A. Harrington for microinjection and significant technical assistance and H. Chen and J. Pinkas for extensive review of this manuscript. We also thank M. Bedford, H. Chen, K Fitzgerald, and J. Pinkas for technical assistance and advice on experimental approaches.

Author information

Author notes

  1. Daniel I. Kitsberg

    Present address: Alomone Labs, POB 4287, Jerusalem, 91042, Israel

Authors and Affiliations

  1. Department of Genetics, Harvard Medical School, Howard Hughes Medical Institute, Boston, Massachusetts, 02115, USA

    Edward J. Weinstein, Daniel I. Kitsberg &  Philip Leder

Authors
  1. Edward J. Weinstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daniel I. Kitsberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Philip Leder
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Philip Leder.

Additional information

Communicated by P. Leder.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Weinstein, E.J., Kitsberg, D.I. & Leder, P. A Mouse Model for Breast Cancer Induced by Amplification and Overexpression of the neu Promoter and Transgene. Mol Med 6, 4–16 (2000). https://doi.org/10.1007/BF03401930

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03401930

Keywords

Search

Navigation