Effects of phosphates on the expression of tissue-nonspecific alkaline phosphatase gene and phosphate-regulating genes in short-term cultures of human osteosarcoma cell lines.
We studied the effects of phosphates on the expression of the human tissue-nonspecific alkaline phosphatase (TNSALP) gene and phosphate-regulating genes in short-term cultures of human osteoblastic osteosarcoma cell lines. When human osteosarcoma cell lines, SaOS-2, MG-63, and U(2)OS were cultured with 10 mM inorganic sodium dihydrogenphosphate, 10 mM beta-glycerophosphate, 250 microM pyridoxal phosphate, or 100 microM inorganic pyrophosphate, enzymatic activity of alkaline phosphatase began to increase at 72 h after addition of sodium dihydrogenphosphate and beta-glycerophosphate in SaOS-2 cells. Pyridoxal phosphate and pyrophosphate did not induce alkaline phosphatase activity. U(2)OS cells slightly reacted to beta-glycerophosphate, but MG-63 cells did not react on exposure to phosphates. In SaOS-2 cells, TNSALP mRNA measured by real-time RT-PCR reached a peak level at 72 h after the addition of beta-glycerophosphate. PHEX and MEPE mRNAs were also induced by beta-glycerophosphate. These results suggest that TNSALP, PHEX and MEPE were concordantly induced by beta-glycerophosphate on mineralisation.
Orimo
Hideo
H
Department of Biochemistry and Molecular Biology, Nippon Medical School, 1-1-5 Sendagi, Tokyo 113-8602, Japan. orimohd@nms.ac.jp
Shimada
Takashi
T
D000469:Alkaline Phosphatase / Q000378:metabolism*; D002113:Calcification, Physiologic; D045744:Cell Line, Tumor; D004789:Enzyme Activation; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D010755:Organophosphates / Q000494:pharmacology*; D012516:Osteosarcoma; D010710:Phosphates / Q000494:pharmacology*; D016226:Phosphorus, Dietary / Q000008:administration & dosage
D010755:Organophosphates; D010710:Phosphates; D016226:Phosphorus, Dietary; D000469:Alkaline Phosphatase