Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1994 Sep;14(9):6278–6286. doi: 10.1128/mcb.14.9.6278

Differential transformation of mammary epithelial cells by Wnt genes.

G T Wong 1, B J Gavin 1, A P McMahon 1
PMCID: PMC359154  PMID: 8065359

Abstract

The mouse Wnt family includes at least 10 genes that encode structurally related secreted glycoproteins. Wnt-1 and Wnt-3 were originally identified as oncogenes activated by the insertion of mouse mammary tumor virus in virus-induced mammary adenocarcinomas, although they are not expressed in the normal mammary gland. However, five other Wnt genes are differentially expressed during development of adult mammary tissue, suggesting that they may play distinct roles in various phases of mammary gland growth and development. Induction of transformation by Wnt-1 and Wnt-3 may be due to interference with these normal regulatory events; however, there is no direct evidence for this hypothesis. We have tested Wnt family members for the ability to induce transformation of cultured mammary cells. The results demonstrate that the Wnt gene family can be divided into three groups depending on their ability to induce morphological transformation and altered growth characteristics of the C57MG mammary epithelial cell line. Wnt-1, Wnt-3A, and Wnt-7A were highly transforming and induced colonies which formed and shed balls of cells. Wnt-2, Wnt-5B, and Wnt-7B also induced transformation but with a lower frequency and an apparent decrease in saturation density. In contrast, Wnt-6 and two other family members which are normally expressed in C57MG cells, Wnt-4 and Wnt-5A, failed to induce transformation. These data demonstrate that the Wnt genes have distinct effects on cell growth and should not be regarded as functionally equivalent.

Full text

PDF
6278

Images in this article

6281Image
on p.6281
6283Image
on p.6283

Selected References

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

  1. Blasband A., Schryver B., Papkoff J. The biochemical properties and transforming potential of human Wnt-2 are similar to Wnt-1. Oncogene. 1992 Jan;7(1):153–161. [PubMed] [Google Scholar]
  2. Bolander F. F., Jr, Nicholas K. R., Van Wyk J. J., Topper Y. J. Insulin is essential for accumulation of casein mRNA in mouse mammary epithelial cells. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5682–5684. doi: 10.1073/pnas.78.9.5682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borellini F., Oka T. Growth control and differentiation in mammary epithelial cells. Environ Health Perspect. 1989 Mar;80:85–99. doi: 10.1289/ehp.898085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradley R. S., Brown A. M. The proto-oncogene int-1 encodes a secreted protein associated with the extracellular matrix. EMBO J. 1990 May;9(5):1569–1575. doi: 10.1002/j.1460-2075.1990.tb08276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown A. M., Wildin R. S., Prendergast T. J., Varmus H. E. A retrovirus vector expressing the putative mammary oncogene int-1 causes partial transformation of a mammary epithelial cell line. Cell. 1986 Sep 26;46(7):1001–1009. doi: 10.1016/0092-8674(86)90699-9. [DOI] [PubMed] [Google Scholar]
  6. Bühler T. A., Dale T. C., Kieback C., Humphreys R. C., Rosen J. M. Localization and quantification of Wnt-2 gene expression in mouse mammary development. Dev Biol. 1993 Jan;155(1):87–96. doi: 10.1006/dbio.1993.1009. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Coleman S., Daniel C. W. Inhibition of mouse mammary ductal morphogenesis and down-regulation of the EGF receptor by epidermal growth factor. Dev Biol. 1990 Feb;137(2):425–433. doi: 10.1016/0012-1606(90)90267-m. [DOI] [PubMed] [Google Scholar]
  9. Daniel C. W., Silberstein G. B., Van Horn K., Strickland P., Robinson S. TGF-beta 1-induced inhibition of mouse mammary ductal growth: developmental specificity and characterization. Dev Biol. 1989 Sep;135(1):20–30. doi: 10.1016/0012-1606(89)90154-1. [DOI] [PubMed] [Google Scholar]
  10. Edwards P. A., Hiby S. E., Papkoff J., Bradbury J. M. Hyperplasia of mouse mammary epithelium induced by expression of the Wnt-1 (int-1) oncogene in reconstituted mammary gland. Oncogene. 1992 Oct;7(10):2041–2051. [PubMed] [Google Scholar]
  11. Gavin B. J., McMahon A. P. Differential regulation of the Wnt gene family during pregnancy and lactation suggests a role in postnatal development of the mammary gland. Mol Cell Biol. 1992 May;12(5):2418–2423. doi: 10.1128/mcb.12.5.2418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gavin B. J., McMahon J. A., McMahon A. P. Expression of multiple novel Wnt-1/int-1-related genes during fetal and adult mouse development. Genes Dev. 1990 Dec;4(12B):2319–2332. doi: 10.1101/gad.4.12b.2319. [DOI] [PubMed] [Google Scholar]
  13. González F., Swales L., Bejsovec A., Skaer H., Martinez Arias A. Secretion and movement of wingless protein in the epidermis of the Drosophila embryo. Mech Dev. 1991 Aug;35(1):43–54. doi: 10.1016/0925-4773(91)90040-d. [DOI] [PubMed] [Google Scholar]
  14. Goodwin L., Hill J. E., Raynor K., Raszi L., Manabe M., Cowin P. Desmoglein shows extensive homology to the cadherin family of cell adhesion molecules. Biochem Biophys Res Commun. 1990 Dec 31;173(3):1224–1230. doi: 10.1016/s0006-291x(05)80917-9. [DOI] [PubMed] [Google Scholar]
  15. Inagaki Y., Kohmoto K. Changes in Scatchard plots for insulin binding to mammary epithelial cells from cycling, pregnant, and lactating mice. Endocrinology. 1982 Jan;110(1):176–182. doi: 10.1210/endo-110-1-176. [DOI] [PubMed] [Google Scholar]
  16. Jhappan C., Stahle C., Harkins R. N., Fausto N., Smith G. H., Merlino G. T. TGF alpha overexpression in transgenic mice induces liver neoplasia and abnormal development of the mammary gland and pancreas. Cell. 1990 Jun 15;61(6):1137–1146. doi: 10.1016/0092-8674(90)90076-q. [DOI] [PubMed] [Google Scholar]
  17. Jue S. F., Bradley R. S., Rudnicki J. A., Varmus H. E., Brown A. M. The mouse Wnt-1 gene can act via a paracrine mechanism in transformation of mammary epithelial cells. Mol Cell Biol. 1992 Jan;12(1):321–328. doi: 10.1128/mcb.12.1.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Klingensmith J., Noll E., Perrimon N. The segment polarity phenotype of Drosophila involves differential tendencies toward transformation and cell death. Dev Biol. 1989 Jul;134(1):130–145. doi: 10.1016/0012-1606(89)90084-5. [DOI] [PubMed] [Google Scholar]
  19. Koch P. J., Walsh M. J., Schmelz M., Goldschmidt M. D., Zimbelmann R., Franke W. W. Identification of desmoglein, a constitutive desmosomal glycoprotein, as a member of the cadherin family of cell adhesion molecules. Eur J Cell Biol. 1990 Oct;53(1):1–12. [PubMed] [Google Scholar]
  20. Korman N. J., Eyre R. W., Klaus-Kovtun V., Stanley J. R. Demonstration of an adhering-junction molecule (plakoglobin) in the autoantigens of pemphigus foliaceus and pemphigus vulgaris. N Engl J Med. 1989 Sep 7;321(10):631–635. doi: 10.1056/NEJM198909073211002. [DOI] [PubMed] [Google Scholar]
  21. Mason J. O., Kitajewski J., Varmus H. E. Mutational analysis of mouse Wnt-1 identifies two temperature-sensitive alleles and attributes of Wnt-1 protein essential for transformation of a mammary cell line. Mol Biol Cell. 1992 May;3(5):521–533. doi: 10.1091/mbc.3.5.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Matsui Y., Halter S. A., Holt J. T., Hogan B. L., Coffey R. J. Development of mammary hyperplasia and neoplasia in MMTV-TGF alpha transgenic mice. Cell. 1990 Jun 15;61(6):1147–1155. doi: 10.1016/0092-8674(90)90077-r. [DOI] [PubMed] [Google Scholar]
  23. McCrea P. D., Turck C. W., Gumbiner B. A homolog of the armadillo protein in Drosophila (plakoglobin) associated with E-cadherin. Science. 1991 Nov 29;254(5036):1359–1361. doi: 10.1126/science.1962194. [DOI] [PubMed] [Google Scholar]
  24. McGrew L. L., Otte A. P., Moon R. T. Analysis of Xwnt-4 in embryos of Xenopus laevis: a Wnt family member expressed in the brain and floor plate. Development. 1992 Jun;115(2):463–473. doi: 10.1242/dev.115.2.463. [DOI] [PubMed] [Google Scholar]
  25. McMahon A. P., Moon R. T. Ectopic expression of the proto-oncogene int-1 in Xenopus embryos leads to duplication of the embryonic axis. Cell. 1989 Sep 22;58(6):1075–1084. doi: 10.1016/0092-8674(89)90506-0. [DOI] [PubMed] [Google Scholar]
  26. Moon R. T., Campbell R. M., Christian J. L., McGrew L. L., Shih J., Fraser S. Xwnt-5A: a maternal Wnt that affects morphogenetic movements after overexpression in embryos of Xenopus laevis. Development. 1993 Sep;119(1):97–111. doi: 10.1242/dev.119.1.97. [DOI] [PubMed] [Google Scholar]
  27. Morata G., Lawrence P. A. The development of wingless, a homeotic mutation of Drosophila. Dev Biol. 1977 Apr;56(2):227–240. doi: 10.1016/0012-1606(77)90266-4. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Nusse R., Varmus H. E. Wnt genes. Cell. 1992 Jun 26;69(7):1073–1087. doi: 10.1016/0092-8674(92)90630-u. [DOI] [PubMed] [Google Scholar]
  30. Nusse R., van Ooyen A., Cox D., Fung Y. K., Varmus H. Mode of proviral activation of a putative mammary oncogene (int-1) on mouse chromosome 15. Nature. 1984 Jan 12;307(5947):131–136. doi: 10.1038/307131a0. [DOI] [PubMed] [Google Scholar]
  31. Nüsslein-Volhard C., Wieschaus E. Mutations affecting segment number and polarity in Drosophila. Nature. 1980 Oct 30;287(5785):795–801. doi: 10.1038/287795a0. [DOI] [PubMed] [Google Scholar]
  32. Oka T., Perry J. W., Topper Y. J. Changes in insulin responsiveness during development of mammary epithelium. J Cell Biol. 1974 Aug;62(2):550–556. doi: 10.1083/jcb.62.2.550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Papkoff J., Schryver B. Secreted int-1 protein is associated with the cell surface. Mol Cell Biol. 1990 Jun;10(6):2723–2730. doi: 10.1128/mcb.10.6.2723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Reynolds A. B., Herbert L., Cleveland J. L., Berg S. T., Gaut J. R. p120, a novel substrate of protein tyrosine kinase receptors and of p60v-src, is related to cadherin-binding factors beta-catenin, plakoglobin and armadillo. Oncogene. 1992 Dec;7(12):2439–2445. [PubMed] [Google Scholar]
  35. 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]
  36. Rijsewijk F., van Deemter L., Wagenaar E., Sonnenberg A., Nusse R. Transfection of the int-1 mammary oncogene in cuboidal RAC mammary cell line results in morphological transformation and tumorigenicity. EMBO J. 1987 Jan;6(1):127–131. doi: 10.1002/j.1460-2075.1987.tb04729.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Sandgren E. P., Luetteke N. C., Palmiter R. D., Brinster R. L., Lee D. C. Overexpression of TGF alpha in transgenic mice: induction of epithelial hyperplasia, pancreatic metaplasia, and carcinoma of the breast. Cell. 1990 Jun 15;61(6):1121–1135. doi: 10.1016/0092-8674(90)90075-p. [DOI] [PubMed] [Google Scholar]
  39. Smith W. C., Harland R. M. Injected Xwnt-8 RNA acts early in Xenopus embryos to promote formation of a vegetal dorsalizing center. Cell. 1991 Nov 15;67(4):753–765. doi: 10.1016/0092-8674(91)90070-f. [DOI] [PubMed] [Google Scholar]
  40. Sokol S., Christian J. L., Moon R. T., Melton D. A. Injected Wnt RNA induces a complete body axis in Xenopus embryos. Cell. 1991 Nov 15;67(4):741–752. doi: 10.1016/0092-8674(91)90069-b. [DOI] [PubMed] [Google Scholar]
  41. Soriano P., Montgomery C., Geske R., Bradley A. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell. 1991 Feb 22;64(4):693–702. doi: 10.1016/0092-8674(91)90499-o. [DOI] [PubMed] [Google Scholar]
  42. Takeichi M. Cadherins: a molecular family important in selective cell-cell adhesion. Annu Rev Biochem. 1990;59:237–252. doi: 10.1146/annurev.bi.59.070190.001321. [DOI] [PubMed] [Google Scholar]
  43. Taketani Y., Oka T. Epidermal growth factor stimulates cell proliferation and inhibits functional differentiation of mouse mammary epithelial cells in culture. Endocrinology. 1983 Sep;113(3):871–877. doi: 10.1210/endo-113-3-871. [DOI] [PubMed] [Google Scholar]
  44. Tonelli Q. J., Sorof S. Epidermal growth factor requirement for development of cultured mammary gland. Nature. 1980 May 22;285(5762):250–252. doi: 10.1038/285250a0. [DOI] [PubMed] [Google Scholar]
  45. Tsukamoto A. S., Grosschedl R., Guzman R. C., Parslow T., Varmus H. E. Expression of the int-1 gene in transgenic mice is associated with mammary gland hyperplasia and adenocarcinomas in male and female mice. Cell. 1988 Nov 18;55(4):619–625. doi: 10.1016/0092-8674(88)90220-6. [DOI] [PubMed] [Google Scholar]
  46. Vaidya A. B., Lasfargues E. Y., Sheffield J. B., Coutinho W. G. Murine mammary tumor virus (MuMTV) infection of an epithelial cell line established from C57BL/6 mouse mammary glands. Virology. 1978 Oct 1;90(1):12–22. doi: 10.1016/0042-6822(78)90328-8. [DOI] [PubMed] [Google Scholar]
  47. Vonderhaar B. K. Local effects of EGF, alpha-TGF, and EGF-like growth factors on lobuloalveolar development of the mouse mammary gland in vivo. J Cell Physiol. 1987 Sep;132(3):581–584. doi: 10.1002/jcp.1041320324. [DOI] [PubMed] [Google Scholar]
  48. Wainwright B. J., Scambler P. J., Stanier P., Watson E. K., Bell G., Wicking C., Estivill X., Courtney M., Boue A., Pedersen P. S. Isolation of a human gene with protein sequence similarity to human and murine int-1 and the Drosophila segment polarity mutant wingless. EMBO J. 1988 Jun;7(6):1743–1748. doi: 10.1002/j.1460-2075.1988.tb03003.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wieschaus E., Riggleman R. Autonomous requirements for the segment polarity gene armadillo during Drosophila embryogenesis. Cell. 1987 Apr 24;49(2):177–184. doi: 10.1016/0092-8674(87)90558-7. [DOI] [PubMed] [Google Scholar]
  50. Wolda S. L., Moody C. J., Moon R. T. Overlapping expression of Xwnt-3A and Xwnt-1 in neural tissue of Xenopus laevis embryos. Dev Biol. 1993 Jan;155(1):46–57. doi: 10.1006/dbio.1993.1005. [DOI] [PubMed] [Google Scholar]
  51. van Ooyen A., Nusse R. Structure and nucleotide sequence of the putative mammary oncogene int-1; proviral insertions leave the protein-encoding domain intact. Cell. 1984 Nov;39(1):233–240. doi: 10.1016/0092-8674(84)90209-5. [DOI] [PubMed] [Google Scholar]
  52. van den Heuvel M., Nusse R., Johnston P., Lawrence P. A. Distribution of the wingless gene product in Drosophila embryos: a protein involved in cell-cell communication. Cell. 1989 Nov 17;59(4):739–749. doi: 10.1016/0092-8674(89)90020-2. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES