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It has long been recognized that the renal proximal tubular epithelium of the hamster is a bona fide estrogen target tissue. The effect of estrogens on the growth of proximal tubule cell explants and dissociated single cells derived from these explant outgrowths has been studied in culture. Renal tubular cells were grown on a PF-HR-9 basement membrane under serum-free chemically defined culture conditions. The cells of tissue explant outgrowths exhibited ultrastructural features typical of proximal tubules including junctional complexes, numerous mitochondria, peroxisomes, and microvilli. At 7-14 days in culture, cell number was enhanced 3-fold in the presence of either 17 beta-estradiol or diethylstilbestrol. Maximal proliferative response was observed at hormone concentrations of 0.6-1 nM. A similar 3-fold increase in cell number was also seen at 1 nM 17 beta-estradiol in subcultured dissociated single tubular cells derived from hamster renal tubular explant outgrowths at 21 days in culture. Neither progesterone, 5 alpha-dihydrotestosterone, nor the inactive diethylstilbestrol metabolite beta-dienestrol elicited this mitogenic effect. Concomitant exposure of tamoxifen at 3-fold molar excess in culture completely abolished the increase in cell number seen with 17 beta-estradiol. Tubular cells obtained from hamster medulla did not exhibit this proliferative response when exposed similarly to 17 beta-estradiol or diethylstilbestrol. The proliferative effect of estrogens on proximal tubular cell growth appears to be species specific since 17 beta-estradiol did not alter the growth of either rat or guinea pig proximal tubules in culture. In addition, at 7-10 days in culture in the presence of 17 beta-estradiol, [3H]thymidine labeling of hamster tubular cells was enhanced 3-fold. A similar increase in mitoses was also observed in cultures containing these potent estrogens during the same time interval of estrogen exposure. These results clearly indicate that estrogens can directly induce primary epithelial cell proliferation at physiologic concentrations and provide strong additional evidence for an important hormonal role in the neoplastic transformation of the hamster kidney.

Specific estrogen-induced cell proliferation of cultured Syrian hamster renal proximal tubular cells in serum-free chemically defined media.