Hormonal regulation of proliferation of granulosa and Leydig cell lines derived from gonadal tumors of transgenic mice expressing the inhibin-alpha subunit promoter/simian virus 40 T-antigen fusion gene.

We have produced a transgenic (TG) mouse model expressing the Simian Virus 40 T-antigen (Tag) gene, driven by a 6-kb fragment of the mouse inhibin-alpha subunit promoter (inh-alpha). The mice develop gonadal tumors with 100% penetrance by the age of 5-8 months, of granulosa cell origin in the ovary, and of Leydig cell origin in the testis. In the present study, we characterized the hormonal regulation of proliferation of two immortalized cell lines, BLT-1, originating from a Leydig cell tumor, and NT-1, originating from a granulosa cell tumor. [3H]-thymidine incorporation in both types of cells was stimulated by activin (> or = 10-30 microg/l), while inhibin had no effect. Transforming growth factor (TGF)-beta, at > or = 0.01 microg/l, stimulated proliferation of the granulosa tumor cells, but no effect was found on the Leydig tumor cells. Progesterone inhibited the proliferation of both cell lines, although the granulosa tumor cells were clearly less sensitive than the Leydig cells to this effect ( > or = 3 micromol/l vs. > 10 nmol/l, respectively). hCG had no effect on the Leydig tumor cell DNA synthesis whereas at high concentration (100 microg/l) it stimulated that of the granulosa cells. We also investigated in BLT-1 and NT-1 cells whether the proliferative changes were related to concomitant changes in Tag expression. In BLT-1 cells, this was stimulated by activin, progesterone and hCG, even though the latter substance did not affect cell proliferation. In contrast, TGF-beta inhibited Tag expression. In NT-1 cells, the expression of Tag was stimulated by activin, while hCG had no effect. In contrast, it was reduced by progesterone, inhibin and TGF-beta. In conclusion, our results indicate that the granulosa and Leydig tumor cells, despite similar mechanism of immortalization, respond differently to several mitotic stimuli. The responses in the level of Tag expression in these cells did not always correlate with the changes observed in cell proliferation, indicating the independence of these two phenomena.