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Transformation by Ras Suppresses Expression of the Neurotrophic Growth Factor Pleiotrophin*

An 18-kDa protein (p18) was detected in lysates and conditioned medium from contact-arrested NIH 3T3 fibroblasts, but was not detected when the cells were transformed by the oncogeneras. Analysis of transformation-defective cell clones generated after mutagenesis of the ras-retroviral vector used to transduce the ras gene showed an inverse correlation between p18 expression and the degree of transformation. p18 expression was high in non-transformed clones, intermediate in a partially transformed clone, undetectable in fully transformed clones, and detectable only at the non-permissive temperature in a clone which was cold-sensitive for ras transformation. In non-transformed cells, p18 expression varied with the degree of confluence. It was almost undetectable in medium from sparse, proliferating cells, but increased as the cells approached confluence and peaked 2–4 days after confluence. Microsequencing of partially purified p18 identified it as the developmentally regulated neurotrophic factor pleiotrophin. In further experiments, pleiotrophin was undetectable or almost undetectable in medium from fully transformed cells expressing the oncogenes v-src, truncated c-raf, activated c-fms, or polyomavirus middle tumor antigen; it was low but easily detectable in medium from SV40 large tumor antigen-expressing cells, which form soft agar colonies but not foci. Thus, pleiotrophin expression in NIH 3T3 cells is associated with quiescence, and suppression of pleiotrophin is related to oncogenic transformation.