Paradoxical effects of phosphate to directly regulate the level of skeletal alkaline phosphatase activity in human osteosarcoma (SaOS-2) cells and inversely regulate the level of skeletal alkaline phosphatase mRNA.
Recent studies indicate that the amount of alkaline phosphatase (ALP) activity in human osteoblast-line cells is proportional to the concentration of phosphate in the culture medium. The current studies were intended to extend those observations and to determine whether the effects of phosphate (and phosphate esters and analogs) to alter the cellular level of ALP activity, in human osteosarcoma SaOS-2 cells, reflected regulation at the level of transcription. Consistent with previous findings, we found direct, time- and dose-dependent correlations between the concentration of phosphate and the amount of ALP activity/mg cell protein (P < 0.05). Surprisingly, we also found a negative correlation between the phosphate concentration in the medium and the level of skeletal ALP mRNA (e.g., r = -0.98, P < 0.01 at 24 hours). As the highest cellular levels of skeletal ALP activity were associated with the lowest levels of ALP mRNA, these data indicated that the phosphate-dependent increase in ALP activity was not mediated by an increase in transcription and, conversely, that the effect of phosphate withdrawal to decrease ALP activity was not mediated by a decrease in transcription.(ABSTRACT TRUNCATED AT 250 WORDS)
Kyeyune-Nyombi
E
E
J. L. Pettis Memorial Veterans Medical Center, Loma Linda University, California 92357, USA.
Nicolas
V
V
Strong
D D
DD
Farley
J
J
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005994:Glycerophosphates / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D012516:Osteosarcoma / Q000201:enzymology* / Q000235:genetics; D010710:Phosphates / Q000008:administration & dosage / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000201:enzymology
D018076:DNA, Complementary; D005994:Glycerophosphates; D010710:Phosphates; D012333:RNA, Messenger; D000469:Alkaline Phosphatase; C031463:beta-glycerophosphoric acid