Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines

Mozeena Bano, Ruth Lupu, Marc E Lippman, Marc E. Lippman, Robert B. Dickson

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A mammary-derived growth factor, MDGF1, which stimulates collagen synthesis and proliferation in mammary epithelial cells was previously detected and purified from human milk and primary human breast tumors. MDGF1 binds to putative cell-surface receptors of 120-140 kDa and stimulates proliferation of normal and malignant human mammary epithelial cells. Partial protein sequence (N-terminal 18 amino acid sequence) shows that MDGF1 has no homology to any other known growth-promoting peptides. Polyclonal antiserum raised against this synthetic peptide recognizes native milk-derived MDGF1. We hypothesize that MDGF1 might be an autocrine or paracrine factor produced by and acting on normal and malignant human breast epithelial cells possessing MDGF1 receptors. As a first step in testing this possibility, we examined whether human breast epithelial cells in culture produce the growth factor. A protein with the size of MDGF1 was immunologically detected in the concentrated conditioned medium prepared from human breast cancer cell line MDA-MB 231, the mammary-derived but nontumorigenic HBL-100 line, and the normal reduction mammoplasty-derived, nonimmortalized 184 cell strain. A competitive radioreceptor assay (RRA) was used to estimate the level of MDGF1 in the conditioned medium. MDGF1 was present in the nanogram range per 1 million cells. A 62-kDa protein was detected in the above cell lysates by Western immunoblotting or by immunoprecipitation of metabolically labeled cell-conditioned media. The polyclonal antisera directed against the 18 amino acid peptide sequence from milk-derived MDGF1 could adsorb MDGF1 biological activity from conditioned medium. In vitro translation of cell mRNA yielded a protein of 55 kDa which was immunoprecipitated by anti-MDGF1 antibody. N-Glycosylation of MDGF1 was suggested by results of experiments using tunicamycin or N-glycanase treatment.

Original languageEnglish
Pages (from-to)610-616
Number of pages7
JournalBiochemistry
Volume31
Issue number2
StatePublished - Dec 1 1992
Externally publishedYes

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Conditioned Culture Medium
Breast
Epithelial Cells
Cell Line
Peptides
Immune Sera
Intercellular Signaling Peptides and Proteins
Proteins
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Glycosylation
Amino Acids
Tunicamycin
Amino Acid Sequence
Milk
Cell Surface Receptors
Bioactivity
Breast Neoplasms
Tumors
Assays
Radioligand Assay

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bano, M., Lupu, R., Lippman, M. E., Lippman, M. E., & Dickson, R. B. (1992). Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines. Biochemistry, 31(2), 610-616.

Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines. / Bano, Mozeena; Lupu, Ruth; Lippman, Marc E; Lippman, Marc E.; Dickson, Robert B.

In: Biochemistry, Vol. 31, No. 2, 01.12.1992, p. 610-616.

Research output: Contribution to journalArticle

Bano, M, Lupu, R, Lippman, ME, Lippman, ME & Dickson, RB 1992, 'Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines', Biochemistry, vol. 31, no. 2, pp. 610-616.
Bano M, Lupu R, Lippman ME, Lippman ME, Dickson RB. Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines. Biochemistry. 1992 Dec 1;31(2):610-616.
Bano, Mozeena ; Lupu, Ruth ; Lippman, Marc E ; Lippman, Marc E. ; Dickson, Robert B. / Production and characterization of mammary-derived growth factor 1 in mammary epithelial cell lines. In: Biochemistry. 1992 ; Vol. 31, No. 2. pp. 610-616.
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