Identification of glycosylated 38-kDa connective tissue growth factor (IGFBP-related protein 2) and proteolytic fragments in human biological fluids, and up-regulation of IGFBP-rP2 expression by TFG-β in Hs578T human breast cancer cells

Doo Hyun Yang, Ho Seong Kim, Elizabeth M. Wilson, Ronald (Ron) Rosenfeld, Youngman Oh

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

Connective Tissue Growth Factor (CTGF) is a cysteine-rich peptide involved in human atherosclerosis and fibrotic disorders such as scleroderma. CTGF has considerable N-terminal sequence similarity with the insulin-like growth factor binding proteins (IGFBPs), including preservation of cysteines, and has been postulated to be a member of the IGFBP superfamily. Indeed, recent studies have shown that baculovirus generated CTGF, a secreted 38-kDa protein, binds IGFs in a specific manner, leading to the provisional renaming of CTGF as IGFBP-8 (or IGFBP-rP2). With immunoprecipitation and immunoblotting, using polyclonal anti-IGFBP-rP2 antibody generated against recombinant human IGFBP-rP2(bac), IGFBP-rP2 can be identified in the serum- free conditioned media of Hs578T human breast cancer cells, as well as in various human biological fluids, such as normal sera, pregnancy sera, and cerebrospinal, amniotic, follicular and peritoneal fluids. Glycosylation studies with endoglycosidase F reveal that endogenous human IGFBP-rP2 is a secreted, glycosylated, approximately 32-38-kDa protein with 2-8-kDa of N- linked sugars and a 30-kDa core. There are 18- and 24-kDa proteins that appear to be IGFBP-rP2 degradation products. In Hs578T human breast cancer cells, transforming growth factor (TGF)-β2, a potent growth inhibitor for these cells, upregulates tGFBP-rP2 mRNA and protein levels. Expression of Hs578T IGFBP-rP2 is significantly increased by TGF-β2 treatment in a dose- dependent manner, with 2.5- and 6-fold increases in mRNA and protein levels, respectively, at a TGF-β2 concentration of 10 ng/ml. Our studies indicate that IGFBP-rP2 appears to be an important endocrine factor, and one of the critical downstream effectors of TGF-β, similar to the role of IGFBP-3 in TGF-β-induced growth inhibition in human breast cancer cells.

Original languageEnglish (US)
Pages (from-to)2593-2596
Number of pages4
JournalJournal of Clinical Endocrinology and Metabolism
Volume83
Issue number7
DOIs
StatePublished - 1998

Fingerprint

Connective Tissue Growth Factor
Insulin-Like Growth Factor Binding Proteins
Up-Regulation
Cells
Breast Neoplasms
Fluids
Transforming Growth Factors
Proteins
Cysteine
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Glycosylation
Follicular Fluid
Growth Inhibitors
Messenger RNA
Insulin-Like Growth Factor Binding Protein 3
Baculoviridae
Ascitic Fluid
Serum-Free Culture Media
Cell growth
Amniotic Fluid

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

Cite this

@article{3c84a70665024731a38f95088fb192e7,
title = "Identification of glycosylated 38-kDa connective tissue growth factor (IGFBP-related protein 2) and proteolytic fragments in human biological fluids, and up-regulation of IGFBP-rP2 expression by TFG-β in Hs578T human breast cancer cells",
abstract = "Connective Tissue Growth Factor (CTGF) is a cysteine-rich peptide involved in human atherosclerosis and fibrotic disorders such as scleroderma. CTGF has considerable N-terminal sequence similarity with the insulin-like growth factor binding proteins (IGFBPs), including preservation of cysteines, and has been postulated to be a member of the IGFBP superfamily. Indeed, recent studies have shown that baculovirus generated CTGF, a secreted 38-kDa protein, binds IGFs in a specific manner, leading to the provisional renaming of CTGF as IGFBP-8 (or IGFBP-rP2). With immunoprecipitation and immunoblotting, using polyclonal anti-IGFBP-rP2 antibody generated against recombinant human IGFBP-rP2(bac), IGFBP-rP2 can be identified in the serum- free conditioned media of Hs578T human breast cancer cells, as well as in various human biological fluids, such as normal sera, pregnancy sera, and cerebrospinal, amniotic, follicular and peritoneal fluids. Glycosylation studies with endoglycosidase F reveal that endogenous human IGFBP-rP2 is a secreted, glycosylated, approximately 32-38-kDa protein with 2-8-kDa of N- linked sugars and a 30-kDa core. There are 18- and 24-kDa proteins that appear to be IGFBP-rP2 degradation products. In Hs578T human breast cancer cells, transforming growth factor (TGF)-β2, a potent growth inhibitor for these cells, upregulates tGFBP-rP2 mRNA and protein levels. Expression of Hs578T IGFBP-rP2 is significantly increased by TGF-β2 treatment in a dose- dependent manner, with 2.5- and 6-fold increases in mRNA and protein levels, respectively, at a TGF-β2 concentration of 10 ng/ml. Our studies indicate that IGFBP-rP2 appears to be an important endocrine factor, and one of the critical downstream effectors of TGF-β, similar to the role of IGFBP-3 in TGF-β-induced growth inhibition in human breast cancer cells.",
author = "Yang, {Doo Hyun} and Kim, {Ho Seong} and Wilson, {Elizabeth M.} and Rosenfeld, {Ronald (Ron)} and Youngman Oh",
year = "1998",
doi = "10.1210/jc.83.7.2593",
language = "English (US)",
volume = "83",
pages = "2593--2596",
journal = "Journal of Clinical Endocrinology and Metabolism",
issn = "0021-972X",
publisher = "The Endocrine Society",
number = "7",

}

TY - JOUR

T1 - Identification of glycosylated 38-kDa connective tissue growth factor (IGFBP-related protein 2) and proteolytic fragments in human biological fluids, and up-regulation of IGFBP-rP2 expression by TFG-β in Hs578T human breast cancer cells

AU - Yang, Doo Hyun

AU - Kim, Ho Seong

AU - Wilson, Elizabeth M.

AU - Rosenfeld, Ronald (Ron)

AU - Oh, Youngman

PY - 1998

Y1 - 1998

N2 - Connective Tissue Growth Factor (CTGF) is a cysteine-rich peptide involved in human atherosclerosis and fibrotic disorders such as scleroderma. CTGF has considerable N-terminal sequence similarity with the insulin-like growth factor binding proteins (IGFBPs), including preservation of cysteines, and has been postulated to be a member of the IGFBP superfamily. Indeed, recent studies have shown that baculovirus generated CTGF, a secreted 38-kDa protein, binds IGFs in a specific manner, leading to the provisional renaming of CTGF as IGFBP-8 (or IGFBP-rP2). With immunoprecipitation and immunoblotting, using polyclonal anti-IGFBP-rP2 antibody generated against recombinant human IGFBP-rP2(bac), IGFBP-rP2 can be identified in the serum- free conditioned media of Hs578T human breast cancer cells, as well as in various human biological fluids, such as normal sera, pregnancy sera, and cerebrospinal, amniotic, follicular and peritoneal fluids. Glycosylation studies with endoglycosidase F reveal that endogenous human IGFBP-rP2 is a secreted, glycosylated, approximately 32-38-kDa protein with 2-8-kDa of N- linked sugars and a 30-kDa core. There are 18- and 24-kDa proteins that appear to be IGFBP-rP2 degradation products. In Hs578T human breast cancer cells, transforming growth factor (TGF)-β2, a potent growth inhibitor for these cells, upregulates tGFBP-rP2 mRNA and protein levels. Expression of Hs578T IGFBP-rP2 is significantly increased by TGF-β2 treatment in a dose- dependent manner, with 2.5- and 6-fold increases in mRNA and protein levels, respectively, at a TGF-β2 concentration of 10 ng/ml. Our studies indicate that IGFBP-rP2 appears to be an important endocrine factor, and one of the critical downstream effectors of TGF-β, similar to the role of IGFBP-3 in TGF-β-induced growth inhibition in human breast cancer cells.

AB - Connective Tissue Growth Factor (CTGF) is a cysteine-rich peptide involved in human atherosclerosis and fibrotic disorders such as scleroderma. CTGF has considerable N-terminal sequence similarity with the insulin-like growth factor binding proteins (IGFBPs), including preservation of cysteines, and has been postulated to be a member of the IGFBP superfamily. Indeed, recent studies have shown that baculovirus generated CTGF, a secreted 38-kDa protein, binds IGFs in a specific manner, leading to the provisional renaming of CTGF as IGFBP-8 (or IGFBP-rP2). With immunoprecipitation and immunoblotting, using polyclonal anti-IGFBP-rP2 antibody generated against recombinant human IGFBP-rP2(bac), IGFBP-rP2 can be identified in the serum- free conditioned media of Hs578T human breast cancer cells, as well as in various human biological fluids, such as normal sera, pregnancy sera, and cerebrospinal, amniotic, follicular and peritoneal fluids. Glycosylation studies with endoglycosidase F reveal that endogenous human IGFBP-rP2 is a secreted, glycosylated, approximately 32-38-kDa protein with 2-8-kDa of N- linked sugars and a 30-kDa core. There are 18- and 24-kDa proteins that appear to be IGFBP-rP2 degradation products. In Hs578T human breast cancer cells, transforming growth factor (TGF)-β2, a potent growth inhibitor for these cells, upregulates tGFBP-rP2 mRNA and protein levels. Expression of Hs578T IGFBP-rP2 is significantly increased by TGF-β2 treatment in a dose- dependent manner, with 2.5- and 6-fold increases in mRNA and protein levels, respectively, at a TGF-β2 concentration of 10 ng/ml. Our studies indicate that IGFBP-rP2 appears to be an important endocrine factor, and one of the critical downstream effectors of TGF-β, similar to the role of IGFBP-3 in TGF-β-induced growth inhibition in human breast cancer cells.

UR - http://www.scopus.com/inward/record.url?scp=0031786081&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031786081&partnerID=8YFLogxK

U2 - 10.1210/jc.83.7.2593

DO - 10.1210/jc.83.7.2593

M3 - Article

VL - 83

SP - 2593

EP - 2596

JO - Journal of Clinical Endocrinology and Metabolism

JF - Journal of Clinical Endocrinology and Metabolism

SN - 0021-972X

IS - 7

ER -