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1 Department of Physiology and 2 Center for Gastrointestinal Biology and Disease, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7545
Paracrine and
autocrine actions of the insulin-like growth factors (IGFs) are
inferred by local expression within the bowel. CCD-18Co cells, IEC-6
cells, and immunoneutralization were used to analyze whether IGFs have
direct autocrine or paracrine effects on proliferation of cultured
intestinal fibroblasts and epithelial cells. Growth factor expression
was analyzed by ribonuclease protection assay and RT-PCR. Extracellular
matrix (ECM) was analyzed for effects on cell proliferation. CCD-18Co
cells express IGF-II mRNAs and low levels of IGF-I mRNA. Conditioned
medium from CCD-18Co cells (CCD-CM) stimulated proliferation of IEC-6
and CCD-18Co cells. Neutralization of IGF immunoreactivity in CCD-CM
reduced but did not abolish this effect. RT-PCR and
immunoneutralization demonstrated that other growth factors contribute
to mitogenic activity of CCD-CM. Preincubation of CCD-CM with ECM
prepared from IEC-6 or CCD-18Co cells reduced its mitogenic activity.
ECM from CCD-18Co cells enhanced growth factor-dependent proliferation of IEC-6 cells. IEC-6 cell ECM inhibited IGF-I action on CCD-18Co cells. We conclude that IGF-II is a potent autocrine mitogen for intestinal fibroblasts. IGF-II interacts with other fibroblast-derived growth factors and ECM to stimulate proliferation of intestinal epithelial cells in a paracrine manner.
intestinal epithelial cells; intestinal fibroblasts; extracellular matrix; conditioned medium
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