Glucose-6-phosphate dehydrogenase activity in monolayer cultures of thyroid epithelial cells: TSH and inhibition of nitrogen oxide synthase affect the enzyme activity and the oxygen sensitivity of the histochemical assay.
The objective was to investigate glucose-6-phosphate dehydrogenase (G6PD) activity in monolayer cultures of thyroid epithelial cells and to examine whether inhibition of nitric oxide synthase affects activity of G6PD or oxygen sensitivity of the assay. Primary cultures without TSH addition prior to experiments demonstrated a TSH-dependent increase in G6PD activity. G6PD activity was higher in F12 medium than in a serum-free physiological medium. Secondary cultures grown in F12 medium demonstrated a diminished activity of G6PD and a lack of response to TSH. In the serum-free physiological medium, G6PD activity was comparable to that found in primary cultures and a response to high concentrations of TSH was maintained. In primary cultures grown in F12 medium devoid of TSH, G6PD activity decreased dose-dependently when nitric oxide synthase activity was inhibited. The oxygen sensitivity of the assay was comparable to that reported previously in malignant cells and correlated with the activity of G6PD in primary cultures. We suggest that thyroid epithelial cells may be an appropriate system to investigate oxygen sensitivity of the G6PD assay as the cells demonstrate a reduced oxygen sensitivity which can be influenced by culture conditions.
Kayser
Lars
L
Department of Medical Anatomy, The Panum Institute, Department of Oto-, Rhino- and Laryngology E, University of Copenhagen, Gentofte County Hospital, Blegdamsvej 3b, Copenhagen N 2200, Denmark. l.kayser@mai.ku.dk
Thomsen
Jens
J
D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D005954:Glucosephosphate Dehydrogenase / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013961:Thyroid Gland / Q000166:cytology*; D013972:Thyrotropin / Q000378:metabolism / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D013972:Thyrotropin; D005954:Glucosephosphate Dehydrogenase; D019001:Nitric Oxide Synthase; D010100:Oxygen