Protein phosphatase 2A (PP2A) acts as a growth suppressor and is negatively influenced by oncogenic signals. We determined its activity in various human breast carcinoma (HBC) cell types to understand its relationship to estrogen receptor (ER) expression as well as to the distribution of protein kinase C (PKC), an opposing enzyme. PP2A activity was measured using a preferred substrate, histone H1 phosphorylated by PKC. PP2A activity was higher in both the soluble and nuclear fractions of ER-positive cell lines (MCF-7, T47D and ZR-75-1) than in the ER-negative cell lines (MDA-MB-231, Hs578T and BT-20). PP2A multiple forms (2A0, 2A1, 2A2), separated by DEAE-cellulose chromatography and immunoblot analysis of PP2A catalytic subunit, also showed similar differences in these two HBC cell types. In all cases, PP2A distribution was inversely correlated with the PKC activity profile. Moreover, PP2A activity in MCF-7 cells maintained in estrogen-depleted medium was low. Nonetheless, it was induced by a prolonged treatment with 17beta-estradiol, this induction being blocked by the antiestrogens, tamoxifen and ICI-182,780. Studies in both MCF-7 transfectants stably overexpressing ras and MDA-MB-231 transfectants stably expressing ER, suggested that a low PP2A distribution in ER-negative HBC cell types may be related to tumor progression rather than the loss of ER. Conceivably, the presence of high PP2A along with low PKC in ER-positive HBC cell types may be related to the restricted cell growth associated with the retention of a certain degree of differentiation or hormonal control. Conversely, the presence of low PP2A along with high PKC in ER-negative cell types may be related to hormone-independent enhanced cell growth.