Apotransferrins from several species promote thyroid hormone-dependent rat pituitary tumor cell growth in iron-restricted serum-free defined culture.

Previously, we have studied thyroid hormone-dependent growth of GH1 rat pituitary tumor cells in iron-restricted serum-free defined medium (Sirbasku, D.A., et al. (1991) Biochemistry 30, 295-304, 7466-7477). Proliferation was promoted by triiodothyronine (T3) and any of seven forms of horse serum-derived apotransferrin (apoTf). In this report, we have asked if apoTfs from other species also acted as thyromedins and if other metal ion chelators served this role. To address these issues, three thyromedins were isolated from human serum and identified as apoTf. Fe3+ depletion, and assay in low-Fe medium, gave ED50s of 1.4-1.7 nM. Fe3+ saturation abolished their activities in high-Fe medium. To ask if apoTf was the major thyromedin in human serum, hormone-depleted preparations were iron saturated and shown to no longer support T3-dependent GH1 cell growth. Next, commercially prepared human, rat, horse, dog, rabbit, guinea pig and mouse apoTfs were shown to be as active under iron-restricted conditions as those isolated from human serum. Bovine apoTf and colostrum lactoferrin were greater than 100-fold less active; human milk apo-lactoferrin and apo-ovotransferrins were inactive. Transferrins which displayed thyromedin activity blocked the binding of 125I-rat 2Fe.Tf to GH1 cell receptors while those without thyromedin activity were ineffective. Finally, the metal ion chelators EDTA, citrate and deferoxamine did not show thyromedin activity indicating that apoTfs uniquely were able to promote T3-dependent cell growth in defined culture.