Autocrine production of prostaglandin F2α enhances phenotypic transformation of normal rat kidney fibroblasts

We have used normal rat kidney (NRK) fibroblasts as an in vitro model system to study cell transformation. These cells obtain a transformed phenotype upon stimulation with growth-modulating factors such as retinoic acid (RA) or transforming growth factor-β (TGF-β). Patch-clamp experiments showed that transformation is paralleled by a profound membrane depolarization from around −70 to −20 mV. This depolarization is caused by a compound in the medium conditioned by transformed NRK cells, which enhances intracellular Ca2+ levels and thereby activates Ca2+-dependent Cl− channels. This compound was identified as prostaglandin F2α (PGF2α) using electrospray ionization mass spectrometry. The active concentration in the medium conditioned by transformed NRK cells as determined using an enzyme immunoassay was 19.7 ± 2.5 nM ( n = 6), compared with 1.5 ± 0.1 nM ( n = 3) conditioned by nontransformed NRK cells. Externally added PGF2α was able to trigger NRK cells that had grown to density arrest to restart their proliferation. This proliferation was inhibited when the FP receptor (i.e., natural receptor for PGF2α) was blocked by AL-8810. RA-induced phenotypic transformation of NRK cells was partially (∼25%) suppressed by AL-8810. Our results demonstrate that PGF2α acts as an autocrine enhancer and paracrine inducer of cell transformation and suggest that it may play a crucial role in carcinogenesis in general.