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. 1995 Mar 14;92(6):2268–2272. doi: 10.1073/pnas.92.6.2268

Insertional mutagenesis identifies a member of the Wnt gene family as a candidate oncogene in the mammary epithelium of int-2/Fgf-3 transgenic mice.

F S Lee 1, T F Lane 1, A Kuo 1, G M Shackleford 1, P Leder 1
PMCID: PMC42465  PMID: 7892260

Abstract

Transgenic mice harboring the int-2/Fgf-3 protooncogene under transcriptional control of the mouse mammary tumor virus (MMTV) promoter/enhancer exhibit a dramatic, benign hyperplasia of the mammary gland. In one int-2 transgenic line (TG.NX), this growth disturbance is evoked by pregnancy and regresses after parturition. Regression of hyperplastic mammary epithelium is less complete after successive pregnancies, and, within 10 months, most TG.NX mice stochastically develop mammary carcinomas that are transplantable in virgin, syngeneic mice. To identify genes that cooperate with int-2 in cell transformation, we infected TG.NX transgenic mice with MMTV. In a cohort of 14 animals, most mammary tumors represented clonal or oligoclonal outgrowths harboring one to five proviral MMTV integrants. Eight of 35 (23%) MMTV+ tumors exhibited proviral insertion at the Wnt-1 locus. No provirus was detected at the int-2, int-3, or Wnt-3 loci. By Southern analysis, two tumors had proviral insertions at the same genomic location, which was mapped to chromosome 15. Cloning of this int locus identified an additional member of the Wnt gene family. The predicted 389-amino acid protein is most closely related to zebrafish Wnt-10a (58% amino acid identity over 362 residues) and, based on homology analysis, was designated Wnt-10b. This newly discovered Wnt family member was expressed in the embryo and mammary gland of virgin but not pregnant mice and represents a candidate collaborating oncogene of int-2/Fgf-3 in the mammary epithelium.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adamson M. C., Dennis C., Delaney S., Christiansen J., Monkley S., Kozak C. A., Wainwright B. Isolation and genetic mapping of two novel members of the murine Wnt gene family, Wnt11 and Wnt12, and the mapping of Wnt5a and Wnt7a. Genomics. 1994 Nov 1;24(1):9–13. doi: 10.1006/geno.1994.1575. [DOI] [PubMed] [Google Scholar]
  2. Christian J. L., Olson D. J., Moon R. T. Xwnt-8 modifies the character of mesoderm induced by bFGF in isolated Xenopus ectoderm. EMBO J. 1992 Jan;11(1):33–41. doi: 10.1002/j.1460-2075.1992.tb05024.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clausse N., Smith R., Calberg-Bacq C. M., Peters G., Dickson C. Mouse mammary-tumor virus activates Fgf-3/Int-2 less frequently in tumors from virgin than from parous mice. Int J Cancer. 1993 Aug 19;55(1):157–163. doi: 10.1002/ijc.2910550128. [DOI] [PubMed] [Google Scholar]
  4. Dixon M., Deed R., Acland P., Moore R., Whyte A., Peters G., Dickson C. Detection and characterization of the fibroblast growth factor-related oncoprotein INT-2. Mol Cell Biol. 1989 Nov;9(11):4896–4902. doi: 10.1128/mcb.9.11.4896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frankel W. N., Stoye J. P., Taylor B. A., Coffin J. M. Genetic identification of endogenous polytropic proviruses by using recombinant inbred mice. J Virol. 1989 Sep;63(9):3810–3821. doi: 10.1128/jvi.63.9.3810-3821.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fung Y. K., Shackleford G. M., Brown A. M., Sanders G. S., Varmus H. E. Nucleotide sequence and expression in vitro of cDNA derived from mRNA of int-1, a provirally activated mouse mammary oncogene. Mol Cell Biol. 1985 Dec;5(12):3337–3344. doi: 10.1128/mcb.5.12.3337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gallahan D., Callahan R. Mammary tumorigenesis in feral mice: identification of a new int locus in mouse mammary tumor virus (Czech II)-induced mammary tumors. J Virol. 1987 Jan;61(1):66–74. doi: 10.1128/jvi.61.1.66-74.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jakobovits A., Shackleford G. M., Varmus H. E., Martin G. R. Two proto-oncogenes implicated in mammary carcinogenesis, int-1 and int-2, are independently regulated during mouse development. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7806–7810. doi: 10.1073/pnas.83.20.7806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kelly G. M., Lai C. J., Moon R. T. Expression of wnt10a in the central nervous system of developing zebrafish. Dev Biol. 1993 Jul;158(1):113–121. doi: 10.1006/dbio.1993.1172. [DOI] [PubMed] [Google Scholar]
  10. Kwan H., Pecenka V., Tsukamoto A., Parslow T. G., Guzman R., Lin T. P., Muller W. J., Lee F. S., Leder P., Varmus H. E. Transgenes expressing the Wnt-1 and int-2 proto-oncogenes cooperate during mammary carcinogenesis in doubly transgenic mice. Mol Cell Biol. 1992 Jan;12(1):147–154. doi: 10.1128/mcb.12.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mansour S. L., Goddard J. M., Capecchi M. R. Mice homozygous for a targeted disruption of the proto-oncogene int-2 have developmental defects in the tail and inner ear. Development. 1993 Jan;117(1):13–28. doi: 10.1242/dev.117.1.13. [DOI] [PubMed] [Google Scholar]
  12. Marchetti A., Robbins J., Campbell G., Buttitta F., Squartini F., Bistocchi M., Callahan R. Host genetic background effect on the frequency of mouse mammary tumor virus-induced rearrangements of the int-1 and int-2 loci in mouse mammary tumors. J Virol. 1991 Aug;65(8):4550–4554. doi: 10.1128/jvi.65.8.4550-4554.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Moore R., Dixon M., Smith R., Peters G., Dickson C. Complete nucleotide sequence of a milk-transmitted mouse mammary tumor virus: two frameshift suppression events are required for translation of gag and pol. J Virol. 1987 Feb;61(2):480–490. doi: 10.1128/jvi.61.2.480-490.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Morris D. W., Barry P. A., Bradshaw H. D., Jr, Cardiff R. D. Insertion mutation of the int-1 and int-2 loci by mouse mammary tumor virus in premalignant and malignant neoplasms from the GR mouse strain. J Virol. 1990 Apr;64(4):1794–1802. doi: 10.1128/jvi.64.4.1794-1802.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Muller W. J., Lee F. S., Dickson C., Peters G., Pattengale P., Leder P. The int-2 gene product acts as an epithelial growth factor in transgenic mice. EMBO J. 1990 Mar;9(3):907–913. doi: 10.1002/j.1460-2075.1990.tb08188.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nusse R., Varmus H. E. Many tumors induced by the mouse mammary tumor virus contain a provirus integrated in the same region of the host genome. Cell. 1982 Nov;31(1):99–109. doi: 10.1016/0092-8674(82)90409-3. [DOI] [PubMed] [Google Scholar]
  17. Ornitz D. M., Moreadith R. W., Leder P. Binary system for regulating transgene expression in mice: targeting int-2 gene expression with yeast GAL4/UAS control elements. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):698–702. doi: 10.1073/pnas.88.3.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Peters G., Brookes S., Smith R., Dickson C. Tumorigenesis by mouse mammary tumor virus: evidence for a common region for provirus integration in mammary tumors. Cell. 1983 Jun;33(2):369–377. doi: 10.1016/0092-8674(83)90418-x. [DOI] [PubMed] [Google Scholar]
  19. Peters G., Lee A. E., Dickson C. Activation of cellular gene by mouse mammary tumour virus may occur early in mammary tumour development. Nature. 1984 May 17;309(5965):273–275. doi: 10.1038/309273a0. [DOI] [PubMed] [Google Scholar]
  20. Peters G., Lee A. E., Dickson C. Concerted activation of two potential proto-oncogenes in carcinomas induced by mouse mammary tumour virus. Nature. 1986 Apr 17;320(6063):628–631. doi: 10.1038/320628a0. [DOI] [PubMed] [Google Scholar]
  21. Rijsewijk F., Schuermann M., Wagenaar E., Parren P., Weigel D., Nusse R. The Drosophila homolog of the mouse mammary oncogene int-1 is identical to the segment polarity gene wingless. Cell. 1987 Aug 14;50(4):649–657. doi: 10.1016/0092-8674(87)90038-9. [DOI] [PubMed] [Google Scholar]
  22. Roelink H., Wagenaar E., Lopes da Silva S., Nusse R. Wnt-3, a gene activated by proviral insertion in mouse mammary tumors, is homologous to int-1/Wnt-1 and is normally expressed in mouse embryos and adult brain. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4519–4523. doi: 10.1073/pnas.87.12.4519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shackleford G. M., MacArthur C. A., Kwan H. C., Varmus H. E. Mouse mammary tumor virus infection accelerates mammary carcinogenesis in Wnt-1 transgenic mice by insertional activation of int-2/Fgf-3 and hst/Fgf-4. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):740–744. doi: 10.1073/pnas.90.2.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shackleford G. M., Varmus H. E. Construction of a clonable, infectious, and tumorigenic mouse mammary tumor virus provirus and a derivative genetic vector. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9655–9659. doi: 10.1073/pnas.85.24.9655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sidow A. Diversification of the Wnt gene family on the ancestral lineage of vertebrates. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5098–5102. doi: 10.1073/pnas.89.11.5098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stamp G., Fantl V., Poulsom R., Jamieson S., Smith R., Peters G., Dickson C. Nonuniform expression of a mouse mammary tumor virus-driven int-2/Fgf-3 transgene in pregnancy-responsive breast tumors. Cell Growth Differ. 1992 Dec;3(12):929–938. [PubMed] [Google Scholar]
  27. Wilkinson D. G., Bhatt S., McMahon A. P. Expression pattern of the FGF-related proto-oncogene int-2 suggests multiple roles in fetal development. Development. 1989 Jan;105(1):131–136. doi: 10.1242/dev.105.1.131. [DOI] [PubMed] [Google Scholar]
  28. Wolda S. L., Moon R. T. Cloning and developmental expression in Xenopus laevis of seven additional members of the Wnt family. Oncogene. 1992 Oct;7(10):1941–1947. [PubMed] [Google Scholar]

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