Expression of insulin-like growth factor-II and insulin-like growth factor binding proteins during Caco-2 cell proliferation and differentiation

Jung H.Y. Park, Mark Corkins, Jon A. Vanderhoof, Nia M. Caruso, Marjorie J. Hrbek, Beverly S. Schaffer, Dorothy H. Slentz, Robert H. McCusker, Richard G. MacDonald

Research output: Contribution to journalArticle

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Abstract

The components of the insulin-like growth factor (IGF) axis and their roles in regulating proliferation and differentiation of the human colon adenocarcinoma cell line, Caco-2, have been investigated. Caco-2 cells proliferated in serum-free medium at 75% the rate observed in medium containing 10% fetal bovine serum. IGF-I (10 nM) increased Caco-2 cell growth in serum-free medium, but not to the rate seen with serum. Multiple IGF-II mRNA species were produced by Caco-2 cells, but IGF-I mRNA was undetectable. Secretion of radioimmunoassayable IGF-II corresponded with steady-state levels of IGF-II mRNA, neither of which was observed to change markedly over the course of 16 days of Caco-2 cell differentiation. Levels of sucrase-isomaltase mRNA, a marker for enterocytic differentiation, increased 12-fold between days 5 and 16 of culture. Northern blotting of total RNA and ligand blot and immunoblot analyses of serum-free conditioned medium revealed that Caco-2 cells produce several IGF binding proteins (IGFBPs), including IGFBP-2, -3, and -4, as well as a 31,000 M(r) species that was not identified. The pattern of IGFBP secretion changed dramatically during Caco-2 cell differentiation: IGFBP-3 and IGFBP-2 increased 8.5-fold and 5-fold, respectively, whereas IGFBP-4 and the 31,000 M species decreased 43% and 90%. Caco-2 cell clones stably transfected with a human IGFBP-4 cDNA construct exhibited a 60% increase in steady-state level of IGFBP-4 mRNA, and secreted twice as much IGFBP-4 protein as controls. Moreover, IGFBP-4-overexpressing cells proliferated at only 25% the rate of control cells in serum-free medium, in conjunction with a 70% increase in expression of sucrase-isomaltase. In summary, these studies indicate that a complex IGF axis is involved in autocrine regulation of Caco-2 cell proliferation and differentiation.

Original languageEnglish (US)
Pages (from-to)396-406
Number of pages11
JournalJournal of cellular physiology
Volume166
Issue number2
DOIs
StatePublished - Feb 1 1996

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Insulin-Like Growth Factor Binding Proteins
Insulin-Like Growth Factor II
Caco-2 Cells
Cell proliferation
Insulin-Like Growth Factor Binding Protein 4
Cell Differentiation
Cell Proliferation
Serum-Free Culture Media
Messenger RNA
Oligo-1,6-Glucosidase
Insulin-Like Growth Factor Binding Protein 2
Sucrase
Insulin-Like Growth Factor Binding Protein 3
Somatomedins
Insulin-Like Growth Factor I
Differentiation Antigens
Cell growth
Conditioned Culture Medium
Serum
Northern Blotting

All Science Journal Classification (ASJC) codes

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Expression of insulin-like growth factor-II and insulin-like growth factor binding proteins during Caco-2 cell proliferation and differentiation. / Park, Jung H.Y.; Corkins, Mark; Vanderhoof, Jon A.; Caruso, Nia M.; Hrbek, Marjorie J.; Schaffer, Beverly S.; Slentz, Dorothy H.; McCusker, Robert H.; MacDonald, Richard G.

In: Journal of cellular physiology, Vol. 166, No. 2, 01.02.1996, p. 396-406.

Research output: Contribution to journalArticle

Park, Jung H.Y. ; Corkins, Mark ; Vanderhoof, Jon A. ; Caruso, Nia M. ; Hrbek, Marjorie J. ; Schaffer, Beverly S. ; Slentz, Dorothy H. ; McCusker, Robert H. ; MacDonald, Richard G. / Expression of insulin-like growth factor-II and insulin-like growth factor binding proteins during Caco-2 cell proliferation and differentiation. In: Journal of cellular physiology. 1996 ; Vol. 166, No. 2. pp. 396-406.
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abstract = "The components of the insulin-like growth factor (IGF) axis and their roles in regulating proliferation and differentiation of the human colon adenocarcinoma cell line, Caco-2, have been investigated. Caco-2 cells proliferated in serum-free medium at 75{\%} the rate observed in medium containing 10{\%} fetal bovine serum. IGF-I (10 nM) increased Caco-2 cell growth in serum-free medium, but not to the rate seen with serum. Multiple IGF-II mRNA species were produced by Caco-2 cells, but IGF-I mRNA was undetectable. Secretion of radioimmunoassayable IGF-II corresponded with steady-state levels of IGF-II mRNA, neither of which was observed to change markedly over the course of 16 days of Caco-2 cell differentiation. Levels of sucrase-isomaltase mRNA, a marker for enterocytic differentiation, increased 12-fold between days 5 and 16 of culture. Northern blotting of total RNA and ligand blot and immunoblot analyses of serum-free conditioned medium revealed that Caco-2 cells produce several IGF binding proteins (IGFBPs), including IGFBP-2, -3, and -4, as well as a 31,000 M(r) species that was not identified. The pattern of IGFBP secretion changed dramatically during Caco-2 cell differentiation: IGFBP-3 and IGFBP-2 increased 8.5-fold and 5-fold, respectively, whereas IGFBP-4 and the 31,000 M species decreased 43{\%} and 90{\%}. Caco-2 cell clones stably transfected with a human IGFBP-4 cDNA construct exhibited a 60{\%} increase in steady-state level of IGFBP-4 mRNA, and secreted twice as much IGFBP-4 protein as controls. Moreover, IGFBP-4-overexpressing cells proliferated at only 25{\%} the rate of control cells in serum-free medium, in conjunction with a 70{\%} increase in expression of sucrase-isomaltase. In summary, these studies indicate that a complex IGF axis is involved in autocrine regulation of Caco-2 cell proliferation and differentiation.",
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AU - Vanderhoof, Jon A.

AU - Caruso, Nia M.

AU - Hrbek, Marjorie J.

AU - Schaffer, Beverly S.

AU - Slentz, Dorothy H.

AU - McCusker, Robert H.

AU - MacDonald, Richard G.

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N2 - The components of the insulin-like growth factor (IGF) axis and their roles in regulating proliferation and differentiation of the human colon adenocarcinoma cell line, Caco-2, have been investigated. Caco-2 cells proliferated in serum-free medium at 75% the rate observed in medium containing 10% fetal bovine serum. IGF-I (10 nM) increased Caco-2 cell growth in serum-free medium, but not to the rate seen with serum. Multiple IGF-II mRNA species were produced by Caco-2 cells, but IGF-I mRNA was undetectable. Secretion of radioimmunoassayable IGF-II corresponded with steady-state levels of IGF-II mRNA, neither of which was observed to change markedly over the course of 16 days of Caco-2 cell differentiation. Levels of sucrase-isomaltase mRNA, a marker for enterocytic differentiation, increased 12-fold between days 5 and 16 of culture. Northern blotting of total RNA and ligand blot and immunoblot analyses of serum-free conditioned medium revealed that Caco-2 cells produce several IGF binding proteins (IGFBPs), including IGFBP-2, -3, and -4, as well as a 31,000 M(r) species that was not identified. The pattern of IGFBP secretion changed dramatically during Caco-2 cell differentiation: IGFBP-3 and IGFBP-2 increased 8.5-fold and 5-fold, respectively, whereas IGFBP-4 and the 31,000 M species decreased 43% and 90%. Caco-2 cell clones stably transfected with a human IGFBP-4 cDNA construct exhibited a 60% increase in steady-state level of IGFBP-4 mRNA, and secreted twice as much IGFBP-4 protein as controls. Moreover, IGFBP-4-overexpressing cells proliferated at only 25% the rate of control cells in serum-free medium, in conjunction with a 70% increase in expression of sucrase-isomaltase. In summary, these studies indicate that a complex IGF axis is involved in autocrine regulation of Caco-2 cell proliferation and differentiation.

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