In several cell types heparin exerts an antiproliferative action; here we report that heparin inhibited the growth of human epidermoid carcinoma A431 cells. Heparin binding to the cell surface was necessary for growth inhibition; binding was influenced by the molecular weight of heparin. Inhibition of A431 cell proliferation was evident in the presence and in the absence of serum, thus indicating that heparin did not act by binding and 'subtracting' nutrients or other serum factors. In confluent A431 cells, EGF induced DNA synthesis, but heparin did not inhibit this effect; consistently, it did not affect inositol lipid turnover triggered by EGF or bradykinin.