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To determine the effect of cell density on vascular endothelial growth factor (VEGF) expression and the mechanism of this effect, four human colon cancer cell lines were grown as sparse or confluent monolayers or as spheroids. VEGF mRNA increased > 2-fold in cells grown as confluent monolayers or spheroids compared with cells grown as sparse monolayers. Semiquantitative reverse transcription-PCR demonstrated a 2-fold increase in the larger VEGF mRNA isoform (189 bp) in confluent cells. Sparse cells grown in conditioned medium from confluent cells demonstrated a > 2-fold increase in VEGF mRNA. These data suggest that VEGF expression may be regulated by an unidentified soluble factor.
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