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Interleukin-6 stimulates cell proliferation of rat pituitary clonal cell lines in vitro
1Department of Obstetrics and Gynecology, Osaka University Medical School, Japan.
We investigated the effect of recombinant human IL-6 (rhIL-6) on cell proliferation using the MtT/E rat pituitary tumor cell line, which was recently established by Inoue et al. This cell line expresses the homeodomain protein Pit-1/GHF 1 and does not produce any significant amount of pituitary hormones, but retains its tumorigenicity by back-transplantation into rats, resulting in production of prolactin. MtT/E cells were seeded into Falcon 24-well plates at a density of 2 x 10(4) cells/well in a cultured medium, containing 10% horse serum and 2.5% fetal bovine, with test drug. After four-days (12 days for the time-course study) incubations, the cells were counted using a hemocytometer. Incubation for 4 days with rhIL-6 caused concentration-dependent stimulation of MtT/E cell growth and [3H]-thymidine incorporation into MtT/E cells. Addition of 20 ng/ml rhIL-6 to the culture medium stimulated MtT/E cell growth in a time-dependent manner, withdrawal of rhIL-6 from the culture medium reduced MtT/E cell growth, and re-addition of rhIL-6 to the culture medium again stimulated MtT/E cell growth. Among the cytokines tested, granulocyte colony-stimulating factor (rh G-CSF) also showed a slight but significant mitogenic activity on the MtT/E cells. Analysis of 125I-rhIL-6 binding to the MtT/E cells indicated a dissociation constant of 0.953 x 10(-9) mmol/l and the presence of 968 binding sites per cell.(ABSTRACT TRUNCATED AT 250 WORDS)
We investigated the effect of recombinant human IL-6 (rhIL-6) on cell proliferation using the MtT/E rat pituitary tumor cell line, which was recently established by Inoue et al. This cell line expresses the homeodomain protein Pit-1/GHF 1 and does not produce any significant amount of pituitary hormones, but retains its tumorigenicity by back-transplantation into rats, resulting in production of prolactin. MtT/E cells were seeded into Falcon 24-well plates at a density of 2 x 10(4) cells/well in a cultured medium, containing 10% horse serum and 2.5% fetal bovine, with test drug. After four-days (12 days for the time-course study) incubations, the cells were counted using a hemocytometer. Incubation for 4 days with rhIL-6 caused concentration-dependent stimulation of MtT/E cell growth and [3H]-thymidine incorporation into MtT/E cells. Addition of 20 ng/ml rhIL-6 to the culture medium stimulated MtT/E cell growth in a time-dependent manner, withdrawal of rhIL-6 from the culture medium reduced MtT/E cell growth, and re-addition of rhIL-6 to the culture medium again stimulated MtT/E cell growth. Among the cytokines tested, granulocyte colony-stimulating factor (rh G-CSF) also showed a slight but significant mitogenic activity on the MtT/E cells. Analysis of 125I-rhIL-6 binding to the MtT/E cells indicated a dissociation constant of 0.953 x 10(-9) mmol/l and the presence of 968 binding sites per cell.(ABSTRACT TRUNCATED AT 250 WORDS)