Phenotype-Associated Changes in the Effects of 1,25-Dihydroxyvitamin D3on Alkaline Phosphatase and Bone GLA-Protein of Rat Osteoblastic Cells*

Abstract

The effects of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on alkaline phosphatase (AP) activity and the synthesis of .gamma.-carboxy-glutamic acid containing protein (BGP) were compared in phenotypically distinct cloned cell lines derived from the osteoblast-like rat osteogenic sarcoma line ROS 17/2-8, 1,25(OH)2D3 stimulated AP activity and BGP synthesis in phenotypes which exhibited relatively low basal AP activity and high basal BGP levels. In contrast 1,25(OH)2D3 inhibited AP activity in phenotypes that exhibited high basal AP activity. The latter cells had undetectable BGP levels and the synthesis of this protein failed to respond to the 1,25(OH)2D3 stimulus. In the cells that responded to 1,25(OH)2D3 with an increase in AP activity the effect of the hormone on AP could be blocked by actinomycin-D. However in the cells that responded to 1,25(OH)2D3 with inhibition of AP the effect of the hormone on AP was not influenced by actinomycin-D. The directly opposite effects of 1,25(OH)2 on the AP activity of the respective clones did not change qualitatively at different stages of culture and could not be accounted by differences in the 1,25(OH)2D3 receptor status nor by different effects of the hormone on cell proliferation. These data raise the possibility that the response of AP to 1,25(OH)2D3 in osteoblastic cells depends on their state of phenotypic differentiation. The stimulatory effect of the hormone in low AP-producing cells might be related to differentiation promoting properties of 1,25(OH)2D3. The inhbitory effect of 1,25(OH)2D3 on AP, unlike the stimulatory effect of the hormone does not appear to be mediated by the classical mechanism of 12,5(OH)2D3 action on the genome and might be associated with dedifferentiated osteoblastic cells.