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 R. 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

ASJC Scopus subject areas

  • 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 R.; 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 R. ; 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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