Growth stimulating activity secreted by human anaplastic thyroid carcinoma cells.
In order to clarify the mechanism of rapid growth of anaplastic thyroid carcinoma, growth stimulating activity produced by the cancer cells in culture was studied. A cell line (HTh7) established from a biopsy specimen of anaplastic thyroid carcinoma was used throughout the study. Growth stimulating activity was determined as an activity to increase 3H-thymidine incorporation in rat thyroid cell line (FRTL5). Conditioned medium of HTh7 cells contained significant growth stimulating activity for FRTL5 cells. The activity was separated into two fractions with heparin agarose gel: heparin-binding and heparin-non-binding. In the medium, the heparin-non-binding activity was much greater than the heparin-binding one. The heparin-non-binding activity was acid stable. It was partially purified with gel filtration in an acidic condition followed by reverse phase HPLC. In gel filtration with a Sephacryl S-200 column, the activity was eluted later than the elution volume of cytochrome c (MW 12400) as several separated peaks. In reverse phase HPLC, however, the activity in these peaks was eluted as a single peak. The retention time of the active peak was almost the same as that of recombinant IGF-I. When measured by specific RIAs, the conditioned media concentrated 20 times contained both 0.35 ng/ml of IGF-I and 5.21 ng/ml of IGF-II. As for the heparin-binding mitogenic activity, when applied to heparin affinity HPLC column and eluted with a linear gradient of NaCl, the activity came out as one major peak with approximately 1.0 M NaCl.(ABSTRACT TRUNCATED AT 250 WORDS)
Tanaka
Y
Y
Third Department of Internal Medicine, University of Tokyo, Japan.
Kubota
K
K
Sasaki
N
N
Takaku
F
F
Uchimura
H
H
D002277:Carcinoma / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D006133:Growth Substances / Q000302:isolation & purification / Q000502:physiology*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D007335:Insulin-Like Growth Factor II / Q000494:pharmacology; D011863:Radioimmunoassay; D013964:Thyroid Neoplasms / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D006133:Growth Substances; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II; D006493:Heparin