In the present study we investigated a panel of human breast cancer cell lines which were sensitive or resistant to the cytotoxic drug doxorubicin, for their abilities to synthesize and bind hyaluronan. We found that MDA-231 and HS-578T cells which express very low amounts of estrogen and progesterone receptors, both synthesized hyaluronan and expressed hyaluronan binding sites on the cell surface as did their doxorubicin-adapted counterparts MDA-231 Dox and HS-578T Dox. The binding was highly specific with a K-d of 0.48x10(-9) M. Most of the hyaluronan binding activity was blocked by mAbs against Hermes-l antigen indicating that the adhesion molecule CD44 is responsible for hyaluronan binding. Only 0.5% of the total amount of labeled hyaluronan added to the cultures was degraded during a period of 16 h. The hormone positive receptor cell lines, MCF-5, Zr-5-1 and Zr-5-1 Dox synthesized only minute amounts of hyaluronan and did not bind hyaluronan or express CD44 receptors. Expression of CD44-related hyaluronan receptors and synthesized hyaluronan may endow hormone receptor negative cells with a highly hydrated environment that facilitates cell motility and invasiveness. The lack of CD44 and thereby the lack of ability to bind hyaluronan in the extracellular matrix may contribute to the non-invasive behavior of hormone positive cell lines.