Additive effects of c-erbB-2, c-Ha-ras, and transforming growth factor-alpha genes on in vitro transformation of human mammary epithelial cells - PubMed

Additive effects of c-erbB-2, c-Ha-ras, and transforming growth factor-alpha genes on in vitro transformation of human mammary epithelial cells

F Ciardiello et al. Mol Carcinog. 1992.

Abstract

MCF-10A cells are a spontaneously immortalized untransformed human mammary epithelial cell line. We have previously shown that overexpression of a human point-mutated c-Ha-ras proto-oncogene, the rat c-neu (c-erbB-2) proto-oncogene, or the human transforming growth factor-alpha (TGF-alpha) gene in MCF-10A cells leads to in vitro transformation of such cells. To ascertain whether the introduction of two of these genes into MCF-10A human mammary epithelial cells induces a completely tumorigenic phenotype, we infected MCF-10A Ha-ras and MCF-10A TGF-alpha cells with a recombinant retroviral vector containing the human c-erbB-2 proto-oncogene and the hygromycin-resistance gene. Ten MCF-10A TGF-alpha/c-erbB-2 (MCF-10A TE) and 10 MCF-10A Ha-ras/c-erbB-2 (MCF-10A HE) hygromycin-resistant clones were randomly selected and expanded into cell lines. MCF-10A TE and MCF-10A HE clones expressed a 10-fold to 40-fold increase in p185 erbB-2 protein levels compared with parental uninfected cells. These cells exhibited a fourfold increase in their growth rate in serum-free medium and showed a strongly reduced mitogenic response to exogenous epidermal growth factor or TGF-alpha compared with MCF-10A cells. Moreover, both MCF-10A TE and MCF-10A HE clones exhibited a fivefold to 20-fold higher cloning efficiency in soft agar than MCF-10A Ha-ras, MCF-10A c-erbB-2, or MCF-10A TGF-alpha clones. However, neither MCF-10A TE nor MCF-10A HE cells were able to grow as tumors in vivo when they were injected into nude mice. These results suggest that c-Ha-ras, c-erbB-2, and TGF-alpha genes have an additive effect on the in vitro transformation of an immortalized human mammary epithelial cell line, but that additional genetic changes such as activation of other proto-oncogenes or inactivation of a tumor suppressor gene may be necessary to elicit a fully tumorigenic phenotype.

Similar articles

• Transforming growth factor-alpha expression is enhanced in human mammary epithelial cells transformed by an activated c-Ha-ras protooncogene but not by the c-neu protooncogene, and overexpression of the transforming growth factor-alpha complementary DNA leads to transformation.

  Ciardiello F, McGeady ML, Kim N, Basolo F, Hynes N, Langton BC, Yokozaki H, Saeki T, Elliott JW, Masui H, et al. Ciardiello F, et al. Cell Growth Differ. 1990 Sep;1(9):407-20. Cell Growth Differ. 1990. PMID: 1981145

• Down-regulation of RI alpha subunit of cAMP-dependent protein kinase induces growth inhibition of human mammary epithelial cells transformed by c-Ha-ras and c-erbB-2 proto-oncogenes.

  Ciardiello F, Pepe S, Bianco C, Baldassarre G, Ruggiero A, Bianco C, Selvam MP, Bianco AR, Tortora G. Ciardiello F, et al. Int J Cancer. 1993 Feb 1;53(3):438-43. doi: 10.1002/ijc.2910530315. Int J Cancer. 1993. PMID: 8094073

• Amphiregulin as an autocrine growth factor for c-Ha-ras- and c-erbB-2-transformed human mammary epithelial cells.

  Normanno N, Selvam MP, Qi CF, Saeki T, Johnson G, Kim N, Ciardiello F, Shoyab M, Plowman G, Brandt R, et al. Normanno N, et al. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2790-4. doi: 10.1073/pnas.91.7.2790. Proc Natl Acad Sci U S A. 1994. PMID: 7908443 Free PMC article.

• Expression of transforming growth factor alpha (TGF alpha) in breast cancer.

  Ciardiello F, Kim N, McGeady ML, Liscia DS, Saeki T, Bianco C, Salomon DS. Ciardiello F, et al. Ann Oncol. 1991 Mar;2(3):169-82. doi: 10.1093/oxfordjournals.annonc.a057897. Ann Oncol. 1991. PMID: 2043488 Review.

• The role of erbB-2 and its ligands in growth control of malignant breast epithelium.

  Lupu R, Dickson RB, Lippman ME. Lupu R, et al. Princess Takamatsu Symp. 1991;22:49-60. Princess Takamatsu Symp. 1991. PMID: 1688228 Review.

Cited by

• Overexpression of fatty acid synthase gene activates HER1/HER2 tyrosine kinase receptors in human breast epithelial cells.

  Vazquez-Martin A, Colomer R, Brunet J, Lupu R, Menendez JA. Vazquez-Martin A, et al. Cell Prolif. 2008 Feb;41(1):59-85. doi: 10.1111/j.1365-2184.2007.00498.x. Cell Prolif. 2008. PMID: 18211286 Free PMC article.

• Defective interleukin six expression and responsiveness in human mammary cells transformed by an adeno 5/SV40 hybrid virus.

  Basolo F, Fiore L, Calvo S, Falcone V, Conaldi PG, Fontanini G, Caligo AM, Merlo G, Gluzman Y, Toniolo A. Basolo F, et al. Br J Cancer. 1996 Jun;73(11):1356-61. doi: 10.1038/bjc.1996.258. Br J Cancer. 1996. PMID: 8645579 Free PMC article.

• Differential interleukin-6/Stat3 signaling as a function of cellular context mediates Ras-induced transformation.

  Leslie K, Gao SP, Berishaj M, Podsypanina K, Ho H, Ivashkiv L, Bromberg J. Leslie K, et al. Breast Cancer Res. 2010;12(5):R80. doi: 10.1186/bcr2725. Epub 2010 Oct 7. Breast Cancer Res. 2010. PMID: 20929542 Free PMC article.

• Defective repression of c-myc in breast cancer cells: A loss at the core of the transforming growth factor beta growth arrest program.

  Chen CR, Kang Y, Massagué J. Chen CR, et al. Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):992-9. doi: 10.1073/pnas.98.3.992. Proc Natl Acad Sci U S A. 2001. PMID: 11158583 Free PMC article.

• The role of transforming growth factor alpha production and ErbB-2 overexpression in induction of tumorigenicity of lung epithelial cells.

  Hamburger AW, Fernandes A, Murakami M, Gerwin BI. Hamburger AW, et al. Br J Cancer. 1998 Apr;77(7):1066-71. doi: 10.1038/bjc.1998.178. Br J Cancer. 1998. PMID: 9569041 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Wiley

• Other Literature Sources

  • The Lens - Patent Citations Database

• Medical

  • MedlinePlus Health Information

• Research Materials

  • NCI CPTC Antibody Characterization Program

• Miscellaneous

  • NCI CPTAC Assay Portal