Insulin-like growth factor binding protein-4 expression is dependent on the carbohydrate in the media in HT-29 cells

HT-29 are colonic carcinoma cells that follow a unique pattern of differentiation dependent on the medium's carbohydrate source. This study compared levels of insulin-like growth factor (IGF)-II and IGF binding protein (IGFBP)-4 when HT-29 cells were grown with standard glucose-containing medium versus galactose-containing (glucose-free) medium. Serum-free media conditioned for 24h were collected at low density, pre-confluence, confluence, and 48-h post-confluence. Ligand blotting of the conditioned galactose medium demonstrated low IGFBP-4 levels until the cells approached confluence, when the levels increased significantly. In standard medium, IGFBP-4 levels increased with increasing cell numbers except for a transient decrease at confluence. Radioimmunoassay showed little change in IGF-II concentrations, although HT-29 cells grown with galactose had lower IGF-II concentrations. HT-29 cells treated with retinoic acid had dose-dependent increases in IGFBP-4 and reduced IGF-II expression. These studies suggest that HT-29 cell differentiation correlates with an increase in IGFBP-4 levels.