Summary Ample evidence confirms that certain cancer cells have the capacity to produce multiple peptides as growth factors and that expression of their receptor may act in tumour cell paracrine and/or autocrine loop mechanisms, either by extracellular release of the growth factor or by the tumour itself. To study the possibility of an autocrine growth mechanism in bladder carcinoma, we investigated the ability of various bladder carcinoma cell lines to proliferate in serum-free medium. A rat bladder carcinoma cell line, BC47, demonstrated exponential and density-dependent growth in serum-free medium. Furthermore, conditioned medium from BC47 cells induced growth-stimulating activity for BC47 cells themselves. Purification and further characterization of this activity was performed by chromatographic methods, SDS-PAGE and N-terminal amino acid analysis. Finally, we have identified that a transferrin-like 70-kDa protein is found to be the main growth-promoting factor in this conditioned medium. In addition, specific antibodies against transferrin and the transferrin-receptor inhibit the in vitro growth of this cell line. Our data suggest that this transferrin-like factor possibly acts as an autocrine growth factor for cancer cells.Keywords: autocrine growth factor; bladder carcinoma; rat bladder carcinoma cell line (BC47); serum-free culture; transferrinThe relatively autonomous nature of malignant cells has been known for many years, i.e. they require fewer exogeneous growth factors for optimal growth than do their counterparts. To explain this phenomenon, it has been suggested that cells could become malignant by the endogeneous production of polypeptide growth factors acting on their producer cells via functional external receptors, allowing phenotypic expression of the peptide by the same cell that produces it. This process has been termed 'autocrine secretion' (Sporn and Todaro, 1980; Sporn and Roberts, 1985). There is now much circumstantial and direct evidence to support the original hypothesis. Many types of tumour cells release polypeptide growth factors into their conditioned medium when grown in cell culture, and these same tumour cells often possess functional receptors for the released peptide.The presence of either serum components or substances such as hormones and growth factors is essential to maintain cells in culture. In analysing the regulatory mechanism of growth factors, the importance of cells that can proliferate in chemically defined, serum-free medium without supplements is increasing, because such cells are likely to synthesize factors with growth-stimulating activity (Messing et al, 1984;Matsuda et al, 1989) that can be isolated easily in the absence of extrinsic growth factors. Recently, it was demonstrated that bladder carcinoma cells were able to secrete a variety of autocrine factors, such as tumour-derived adhesion factor (Akaogi et al, 1994) and granulocyte colony-stimulating factor (Tachibana et al, 1995). The present study was undertaken to investigate the ability of various bla...