Induction (or Stimulation) of Prolactin and Growth Hormone Production in a Rat Pituitary Tumor Cell Line by Bromodeoxyuridine

Under basal conditions, a rat pituitary tumor cell line (C/sub 3/ 11RAP) does not secrete any detectable PRL, FSH, and LH, and secretes only minute amounts of GH (27.1 +/- 0.5 ng/10/sup 6/ cells . 24 h), as evaluated by RIA. Bromodeoxyuridine (BrdUrd) added to the culture medium induced the accumulation of PRL into cells and medium, increased that of GH, but did not induce that of LH or FSH. The amount of radioimmunoassayable PRL and GH accumulated in the medium, increased after a lag period of 15 days and was drug concentration dependent. Maximal accumulation was 232.9 +/- 36.8 and 493.6 +/- 41.5 ng/10/sup 6/ cells . 24 h for PRL and GH, respectively, at 50 ..mu..g/ml BrdUrd. In the presence of BrdUrd (greater than or equal to20 ..mu..g/ml), the cells grew more slowly and were more strongly attached to the flasks. All of the effects induced by BrdUrd were reversible. PRL and GH were characterized by three methods: 1) radiocompetition with increasing dilution of samples; 2) Sephadex chromatography, followed by RIAs; and 3) sodium dodecyl sulfate-polyacrylamide gel electrophoresis done on the immunoprecipitate of the proteins secreted by cells incubated with (/sup 3/H)leucine. Chronic treatment with TRH (3 xmore » 10/sup -6/ M) of cells grown without BrdUrd was unable to increase the production of GH or to induce that of PRL. On the other hand, after the same treatment of cells cultured in the presence of BrdUrd, the amounts of PRL accumulated in the culture medium or cells were increased 2- to 7-fold over unstimulated levels; under the same conditions, GH accumulation in the medium was also increased, but this augmentation was less than that of PRL.« less