Demonstration of receptors for insulin-like growth factor binding protein-3 on Hs578T human breast cancer cells

Youngman Oh, Hermann L. Müller, Hung Pham, Ronald (Ron) Rosenfeld

Research output: Contribution to journalArticle

288 Citations (Scopus)

Abstract

Hs578T human breast cancer cells are from an estrogen receptor-negative breast cell line derived from a highly aggressive mammary tumor. Our previous insulin-like growth factor binding protein-3 (IGFBP-S) binding studies (Oh, Y., Müller, H. L., Lamson, G., and Rosenfeld, R. G. (1993) J. Biol. Chem. 268, 14964-14971) have demonstrated specific binding of IGFBP-3 on the Hs578T cell surface and a significant inhibitory effect of IGFBP-3, itself, on monolayer growth. In this study, we have demonstrated cell surface association proteins that are specific for IGFBP-3 by showing: 1) detection of 20-, 26-, and 50-kDa proteins by affinity cross-linking with 125I-IGFBP-3E. coli and immunoprecipitation of cell monolayers and cell lysates with anti-IGFBP-3 antibodies; 2) dose-dependent competition of 125I-IGFBP-3E. coli by unlabeled IGFBP-3E. coli; 3) inhibition of IGFBP-3 binding to these cell surface proteins by EDTA and by coincubation with native insulin-like growth factor II (IGF-II), but not by coincubation with [Gln6,Ala7,Tyr18, Leu19,Leu27]IGF-II, an IGF-II analog with decreased affinity for IGFBP-3; and 4) partial purification of 20- and 26-kDa species by IGFBP-3-anti-IGFBP-3 antibody immunoaffmity membranes. Characteristics of these specific IGFBP-3 cell surface association proteins are identical to those observed in our previous monolayer binding assay and monolayer growth assay experiments. The specificity of binding and the inhibitory effect of IGFBP-3 binding on Hs578T cell growth suggest that these cell surface proteins are IGFBP-3-specific receptors or receptor subunits mediating the direct inhibitory effect of IGFBP-3 on monolayer growth of Hs578T cells.

Original languageEnglish (US)
Pages (from-to)26045-26048
Number of pages4
JournalJournal of Biological Chemistry
Volume268
Issue number35
StatePublished - Dec 15 1993
Externally publishedYes

Fingerprint

Insulin-Like Growth Factor Binding Protein 3
Demonstrations
Cells
Breast Neoplasms
Insulin-Like Growth Factor Binding Proteins
Monolayers
Insulin-Like Growth Factor II
Membrane Proteins
Growth
Assays
Insulin-Like Growth Factor Binding Protein 4
Association reactions
Proteins
Antibodies
Cell growth
Immunoprecipitation
Edetic Acid
Estrogen Receptors
Purification
Tumors

ASJC Scopus subject areas

  • Biochemistry

Cite this

Demonstration of receptors for insulin-like growth factor binding protein-3 on Hs578T human breast cancer cells. / Oh, Youngman; Müller, Hermann L.; Pham, Hung; Rosenfeld, Ronald (Ron).

In: Journal of Biological Chemistry, Vol. 268, No. 35, 15.12.1993, p. 26045-26048.

Research output: Contribution to journalArticle

Oh, Youngman ; Müller, Hermann L. ; Pham, Hung ; Rosenfeld, Ronald (Ron). / Demonstration of receptors for insulin-like growth factor binding protein-3 on Hs578T human breast cancer cells. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 35. pp. 26045-26048.
@article{965060d6a56849d2b1a414e2cceb8024,
title = "Demonstration of receptors for insulin-like growth factor binding protein-3 on Hs578T human breast cancer cells",
abstract = "Hs578T human breast cancer cells are from an estrogen receptor-negative breast cell line derived from a highly aggressive mammary tumor. Our previous insulin-like growth factor binding protein-3 (IGFBP-S) binding studies (Oh, Y., M{\"u}ller, H. L., Lamson, G., and Rosenfeld, R. G. (1993) J. Biol. Chem. 268, 14964-14971) have demonstrated specific binding of IGFBP-3 on the Hs578T cell surface and a significant inhibitory effect of IGFBP-3, itself, on monolayer growth. In this study, we have demonstrated cell surface association proteins that are specific for IGFBP-3 by showing: 1) detection of 20-, 26-, and 50-kDa proteins by affinity cross-linking with 125I-IGFBP-3E. coli and immunoprecipitation of cell monolayers and cell lysates with anti-IGFBP-3 antibodies; 2) dose-dependent competition of 125I-IGFBP-3E. coli by unlabeled IGFBP-3E. coli; 3) inhibition of IGFBP-3 binding to these cell surface proteins by EDTA and by coincubation with native insulin-like growth factor II (IGF-II), but not by coincubation with [Gln6,Ala7,Tyr18, Leu19,Leu27]IGF-II, an IGF-II analog with decreased affinity for IGFBP-3; and 4) partial purification of 20- and 26-kDa species by IGFBP-3-anti-IGFBP-3 antibody immunoaffmity membranes. Characteristics of these specific IGFBP-3 cell surface association proteins are identical to those observed in our previous monolayer binding assay and monolayer growth assay experiments. The specificity of binding and the inhibitory effect of IGFBP-3 binding on Hs578T cell growth suggest that these cell surface proteins are IGFBP-3-specific receptors or receptor subunits mediating the direct inhibitory effect of IGFBP-3 on monolayer growth of Hs578T cells.",
author = "Youngman Oh and M{\"u}ller, {Hermann L.} and Hung Pham and Rosenfeld, {Ronald (Ron)}",
year = "1993",
month = "12",
day = "15",
language = "English (US)",
volume = "268",
pages = "26045--26048",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "35",

}

TY - JOUR

T1 - Demonstration of receptors for insulin-like growth factor binding protein-3 on Hs578T human breast cancer cells

AU - Oh, Youngman

AU - Müller, Hermann L.

AU - Pham, Hung

AU - Rosenfeld, Ronald (Ron)

PY - 1993/12/15

Y1 - 1993/12/15

N2 - Hs578T human breast cancer cells are from an estrogen receptor-negative breast cell line derived from a highly aggressive mammary tumor. Our previous insulin-like growth factor binding protein-3 (IGFBP-S) binding studies (Oh, Y., Müller, H. L., Lamson, G., and Rosenfeld, R. G. (1993) J. Biol. Chem. 268, 14964-14971) have demonstrated specific binding of IGFBP-3 on the Hs578T cell surface and a significant inhibitory effect of IGFBP-3, itself, on monolayer growth. In this study, we have demonstrated cell surface association proteins that are specific for IGFBP-3 by showing: 1) detection of 20-, 26-, and 50-kDa proteins by affinity cross-linking with 125I-IGFBP-3E. coli and immunoprecipitation of cell monolayers and cell lysates with anti-IGFBP-3 antibodies; 2) dose-dependent competition of 125I-IGFBP-3E. coli by unlabeled IGFBP-3E. coli; 3) inhibition of IGFBP-3 binding to these cell surface proteins by EDTA and by coincubation with native insulin-like growth factor II (IGF-II), but not by coincubation with [Gln6,Ala7,Tyr18, Leu19,Leu27]IGF-II, an IGF-II analog with decreased affinity for IGFBP-3; and 4) partial purification of 20- and 26-kDa species by IGFBP-3-anti-IGFBP-3 antibody immunoaffmity membranes. Characteristics of these specific IGFBP-3 cell surface association proteins are identical to those observed in our previous monolayer binding assay and monolayer growth assay experiments. The specificity of binding and the inhibitory effect of IGFBP-3 binding on Hs578T cell growth suggest that these cell surface proteins are IGFBP-3-specific receptors or receptor subunits mediating the direct inhibitory effect of IGFBP-3 on monolayer growth of Hs578T cells.

AB - Hs578T human breast cancer cells are from an estrogen receptor-negative breast cell line derived from a highly aggressive mammary tumor. Our previous insulin-like growth factor binding protein-3 (IGFBP-S) binding studies (Oh, Y., Müller, H. L., Lamson, G., and Rosenfeld, R. G. (1993) J. Biol. Chem. 268, 14964-14971) have demonstrated specific binding of IGFBP-3 on the Hs578T cell surface and a significant inhibitory effect of IGFBP-3, itself, on monolayer growth. In this study, we have demonstrated cell surface association proteins that are specific for IGFBP-3 by showing: 1) detection of 20-, 26-, and 50-kDa proteins by affinity cross-linking with 125I-IGFBP-3E. coli and immunoprecipitation of cell monolayers and cell lysates with anti-IGFBP-3 antibodies; 2) dose-dependent competition of 125I-IGFBP-3E. coli by unlabeled IGFBP-3E. coli; 3) inhibition of IGFBP-3 binding to these cell surface proteins by EDTA and by coincubation with native insulin-like growth factor II (IGF-II), but not by coincubation with [Gln6,Ala7,Tyr18, Leu19,Leu27]IGF-II, an IGF-II analog with decreased affinity for IGFBP-3; and 4) partial purification of 20- and 26-kDa species by IGFBP-3-anti-IGFBP-3 antibody immunoaffmity membranes. Characteristics of these specific IGFBP-3 cell surface association proteins are identical to those observed in our previous monolayer binding assay and monolayer growth assay experiments. The specificity of binding and the inhibitory effect of IGFBP-3 binding on Hs578T cell growth suggest that these cell surface proteins are IGFBP-3-specific receptors or receptor subunits mediating the direct inhibitory effect of IGFBP-3 on monolayer growth of Hs578T cells.

UR - http://www.scopus.com/inward/record.url?scp=0027373968&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027373968&partnerID=8YFLogxK

M3 - Article

VL - 268

SP - 26045

EP - 26048

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 35

ER -