Transformation of an established mouse mammary epithelial cell line following transfection with a human transforming growth factor alpha cDNA

V. Shankar, F. Ciardiello, N. Kim, R. Derynck, D. S. Liscia, G. Merlo, B. C. Langton, D. Sheer, R. Callahan, R. H. Bassin, Marc E Lippman, N. Hynes, D. S. Salomon

Research output: Contribution to journalArticle

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Abstract

To determine whether the enhanced expression of transforming growth factor α (TGFα) is sufficient to induce the neoplastic transformation of an immortalized population of mammary epithelial cells, we cotransfected NOG-8 cells, a cloned mouse mammary epithelial cell line, with a simian virus 40-human TGFα cDNA expression vector plasmid and a pSV2neo plasmid. After cotransfection, nine G418-resistant NOG-8 colonies were cloned and expanded. All clones were subsequently analyzed for TGFα mRNA expression by northern blot analysis, TGFα secretion, anchorage-dependent growth in serum-free medium, anchorage-dependent growth in soft agar, and tumorigenicity in nude mice. Three TGFα-transfected NOG-8 clones expressed high levels of a specific TGFα mRNA, secreted elevated levels of TGFα into the culture medium (177-595 ng/108 cells/48 h), exhibited an enhanced growth rate, grew aggressively as colonies in soft agar, and formed undifferentiated, invasive carcinomas in nude mice. A neutralizing mouse monoclonal antibody generated against the low molecular weight human TGFα peptide was able to inhibit colony formation in soft agar TGFα-transfected NOG-8 clones that produced high levels by TGFα. This inhibition suggested that TGFα acted through an external autocrine loop. NOG-8 cells and NOG-8 cells transfected with a pSV2neo plasmid alone secreted very low levels of TGFα, failed to grow as colonies in soft agar and did not form tumors in nude mice. These results demonstrate that overexpression of a human TGFα cDNA in immortalized, nontransformed mouse mammary epithelial cells can induce a transformed phenotype in vitro and can facilitate tumor formation in vivo.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalMolecular Carcinogenesis
Volume2
Issue number1
StatePublished - Jan 1 1989
Externally publishedYes

Fingerprint

Transforming Growth Factors
Transfection
Breast
Complementary DNA
Epithelial Cells
Cell Line
Agar
Nude Mice
Plasmids
Clone Cells
human TGFA protein
Growth
Messenger RNA
Simian virus 40
Serum-Free Culture Media
Northern Blotting
Culture Media
Neoplasms
Molecular Weight
Monoclonal Antibodies

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology

Cite this

Shankar, V., Ciardiello, F., Kim, N., Derynck, R., Liscia, D. S., Merlo, G., ... Salomon, D. S. (1989). Transformation of an established mouse mammary epithelial cell line following transfection with a human transforming growth factor alpha cDNA. Molecular Carcinogenesis, 2(1), 1-11.

Transformation of an established mouse mammary epithelial cell line following transfection with a human transforming growth factor alpha cDNA. / Shankar, V.; Ciardiello, F.; Kim, N.; Derynck, R.; Liscia, D. S.; Merlo, G.; Langton, B. C.; Sheer, D.; Callahan, R.; Bassin, R. H.; Lippman, Marc E; Hynes, N.; Salomon, D. S.

In: Molecular Carcinogenesis, Vol. 2, No. 1, 01.01.1989, p. 1-11.

Research output: Contribution to journalArticle

Shankar, V, Ciardiello, F, Kim, N, Derynck, R, Liscia, DS, Merlo, G, Langton, BC, Sheer, D, Callahan, R, Bassin, RH, Lippman, ME, Hynes, N & Salomon, DS 1989, 'Transformation of an established mouse mammary epithelial cell line following transfection with a human transforming growth factor alpha cDNA', Molecular Carcinogenesis, vol. 2, no. 1, pp. 1-11.
Shankar, V. ; Ciardiello, F. ; Kim, N. ; Derynck, R. ; Liscia, D. S. ; Merlo, G. ; Langton, B. C. ; Sheer, D. ; Callahan, R. ; Bassin, R. H. ; Lippman, Marc E ; Hynes, N. ; Salomon, D. S. / Transformation of an established mouse mammary epithelial cell line following transfection with a human transforming growth factor alpha cDNA. In: Molecular Carcinogenesis. 1989 ; Vol. 2, No. 1. pp. 1-11.
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