title: 1,25-Dihydroxyvitamin D3 exerts opposite effects on the regulation of human embryonic and nonembryonic alkaline phosphatase isoenzymes.
abstract: The regulation of alkaline phosphatase activity by steroid hormones was studied in two human breast cancer cell lines, MDA-MB-157 and BT20. MDA-MB-157 cells were shown to express the alkaline phosphatase isoenzyme produced by normal breast tissue, and the activity of this isoenzyme increased 3-fold after a 72-h treatment of these cells with 10(-7) M 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3], 2-fold after treatment with 10(-6) M hydrocortisone (HC), and 5-fold after treatment with both hormones. BT20 cells did not express the isoenzyme phenotypic to breast, but ectopically expressed the isoenzyme phenotypic to term placenta and other embryonic tissue. Treatment of BT20 cells with 1,25-(OH)2D3 results in a 30% decrease in alkaline phosphatase activity of the embryonic isoenzyme. There was a 2-fold increase in activity after treatment with HC, and enzyme activity was similar to control values after treatment with both hormones. For both cell lines, changes in alkaline phosphatase activity correlated with changes in nanograms of isoenzyme per mg cellular protein, as measured by RIA. Increases in enzyme activity were inhibited when the cells were incubated simultaneously with the steroids and cycloheximide. Studies with receptors in each cell line showed that both cell lines bound 1,25-(OH)2D3 and that a 1,25-(OH)2D3-binding protein with the same mol wt as the D3 receptor was present in both. The BT20 cells also express a larger mol wt protein which binds 1,25-(OH)2D3 but is not as specific for the 1,25-(OH)2D3 isomer. HC receptors were similar in quantity and binding affinity in both cell lines.
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D001943:Breast Neoplasms / Q000201:enzymology; D002117:Calcitriol / Q000502:physiology*; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism*; D011863:Radioimmunoassay; D018167:Receptors, Calcitriol; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D018167:Receptors, Calcitriol; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D003907:Dexamethasone; D003513:Cycloheximide; D000469:Alkaline Phosphatase; D002117:Calcitriol; D006854:Hydrocortisone