We have established and characterised a cell line, designated WART, from a patient with primary adenocarcinoma of the lung. This cell line grows with a doubling time of approximately 15 hours, forms colonies in soft agarose, is tumorigenic in athymic nude mice, and has a complex karyotype with both structural and numerical abnormalities. WART serum free conditioned medium (SFCM) contains a factor which stimulates motile behavior of WART cells. This factor with an apparent molecular weight of 67 kDa induced in an autocrine fashion prominent pseudopodia, and chemotactic and chemokinetic responses. Heparin affinity chromatography, ion exchange and molecular sieve chromatography accompanied by SDS-PAGE analysis showed that the motility inducing activity was associated with a major band with molecular weight 67 kDa. The motility inducing activity of the 67 kDa protein was not sensitive to reduction with either dithiotreitol or mercaptoethanol which distinguishes it from A-2058 melanoma autocrine motility factor (AMF)/autotaxin, HT-1080 fibrosarcoma AMF and scatter factor which lose their biological activity upon reduction. This 67 kDa motility inducing factor did not augment DNA synthesis indicating that its locomotor activity is independent of mechanisms regulating cell growth. Pertusis toxin inhibited the motile response induced by the 67 kDa protein indicating a signal transduction pathway involving G proteins. Due to its production of the motility stimulating protein the cell line could facilitate studies of invasion and metastasis of human lung tumors.