Autocrine production of prostaglandin F2alpha 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-beta (TGF-beta). 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 F2alpha (PGF2alpha) 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 PGF2alpha 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 PGF2alpha) was blocked by AL-8810. RA-induced phenotypic transformation of NRK cells was partially (approximately 25%) suppressed by AL-8810. Our results demonstrate that PGF2alpha acts as an autocrine enhancer and paracrine inducer of cell transformation and suggest that it may play a crucial role in carcinogenesis in general.
Harks
E G A
EG
Department of Cell Biology, Radboud University Nijmegen, Toernooiveld 1, 6525 ED Nijmegen, The Netherlands.
Peters
P H J
PH
van Dongen
J L J
JL
van Zoelen
E J J
EJ
Theuvenet
A P R
AP
D000818:Animals; D019898:Autocrine Communication / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D002461:Cell Line, Transformed; D017077:Culture Media, Conditioned / Q000494:pharmacology; D015237:Dinoprost / Q000096:biosynthesis*; D004594:Electrophysiology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D007124:Immunoenzyme Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D007668:Kidney / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D013058:Mass Spectrometry; D008564:Membrane Potentials; D009994:Osmolar Concentration; D010641:Phenotype; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D051381:Rats
D017077:Culture Media, Conditioned; D015237:Dinoprost; D011451:Prostaglandin-Endoperoxide Synthases; D002118:Calcium