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. 1979 Oct 1;183(1):73–81. doi: 10.1042/bj1830073

Increase in alkaline phosphatase activity in calvaria cells cultured with diphosphonates.

R Felix, H Fleisch
PMCID: PMC1161474  PMID: 534490

Abstract

1. Dichloromethanediphosphonate and to a lesser degree 1-hydroxyethane-1,1-diphosphonate, two compounds characterized by a P-C-P bond, increased the alkaline phosphatase activity of cultured rat calvaria cells up to 30 times in a dose-dependent fashion. 2. Both diphosphonates also slightly inhibited the protein synthesis in these cells. 3. Thymidine, an inhibitor of cell division, did not inhibit the induction of the enzyme, indicating that the increase in enzyme activity was not due to the formation of a specific population of cells with high alkaline phosphatase activity. 4. The effect on alkaline phosphatase was suppressed by the addition of cycloheximide, an inhibitor of protein synthesis. 5. After subculturing the stimulated cells in medium without diphosphonates, the enzyme activity fell almost to the control value. 6. Bovine parathyrin diminished the enzyme activity of the control cells and the cells treated with dichloromethanediphosphonate; however, at high concentration the effect of parathyrin was greater on the diphosphonate-treated cells than on the control cells. 7. The electrophoretic behaviour, heat inactivation, inhibition by bromotetramisole or by phenylalanine, and the Km value of the induced enzyme were identical with that of the control enzyme.

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Selected References

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  1. Bazzell K. L., Price G., Tu S., Griffin M. Cortisol modification of HeLa 65 alkaline phosphatase. Decreased phosphate content of the induced enzyme. Eur J Biochem. 1976 Jan 15;61(2):493–499. doi: 10.1111/j.1432-1033.1976.tb10044.x. [DOI] [PubMed] [Google Scholar]
  2. Borgers M., Thoné F. The inhibition of alkaline phosphatase by L-p-bromotetramisole. Histochemistry. 1975 Aug 28;44(3):277–280. doi: 10.1007/BF00491496. [DOI] [PubMed] [Google Scholar]
  3. COX R. P., MACLEOD C. M. Hormonal induction of alkaline phosphatase in human cells in tissue culture. Nature. 1961 Apr 1;190:85–87. doi: 10.1038/190085a0. [DOI] [PubMed] [Google Scholar]
  4. COX R. P., PONTECORVO G. Induction of alkaline phosphatase by substrates in established cultures of cells from individual human donors. Proc Natl Acad Sci U S A. 1961 Jun 15;47:839–845. doi: 10.1073/pnas.47.6.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chou J. Y., Robinson J. C. Induction of alkaline phosphatase in choriocarcinoma cells by 1-beta-D-arabinofuranosyl-cytosine, mitomycin C, phleomycin, and cyclic nucleotides. J Cell Physiol. 1977 Aug;92(2):221–231. doi: 10.1002/jcp.1040920210. [DOI] [PubMed] [Google Scholar]
  6. Chou J. Y., Robinson J. C. Induction of placental alkaline phosphatase in choriocarcinoma cells by 5-bromo-2'-deoxyuridine. In Vitro. 1977 Jul;13(7):450–460. doi: 10.1007/BF02615106. [DOI] [PubMed] [Google Scholar]
  7. Cox R. P., Elson N. A., Tu S. H., Griffin M. J. Hormonal induction of alkaline phosphatase activity by an increase in catalytic efficiency of the enzyme. J Mol Biol. 1971 May 28;58(1):197–215. doi: 10.1016/0022-2836(71)90241-5. [DOI] [PubMed] [Google Scholar]
  8. Deutsch S. I., Ghosh N. K., Cox R. P. Increased alkaline phosphatase activity in HeLa cells mediated by aliphatic monocarboxylates and inhibitors of DNA synthesis. Biochim Biophys Acta. 1977 Oct 25;499(3):382–391. doi: 10.1016/0304-4165(77)90069-1. [DOI] [PubMed] [Google Scholar]
  9. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  10. Fast D. K., Felix R., Dowse C., Neuman W. F., Fleisch H. The effects of diphosphonates on the growth and glycolysis of connective-tissue cells in culture. Biochem J. 1978 Apr 15;172(1):97–107. doi: 10.1042/bj1720097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Felix R., Fleisch H. Pyrophosphatase and ATPase of isolated cartilage matrix vesicles. Calcif Tissue Res. 1976 Nov 24;22(1):1–7. doi: 10.1007/BF02010340. [DOI] [PubMed] [Google Scholar]
  12. Felix R., Fleisch H. The pyrophosphatase and (Ca2+-Mg2+)-ATPase activity of purified calf bone alkaline phosphatase. Biochim Biophys Acta. 1974 May 20;350(1):84–94. doi: 10.1016/0005-2744(74)90205-8. [DOI] [PubMed] [Google Scholar]
  13. Felix R., Herrmann W., Fleisch H. Stimulation of precipitation of calcium phosphate by matrix vesicles. Biochem J. 1978 Mar 15;170(3):681–691. doi: 10.1042/bj1700681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fernley H. N., Bisaz S. Studies on alkaline phosphatase. Phosphorylation of calf-intestinal alkaline phosphatase by 32P-labelled pyrophosphate. Biochem J. 1968 Mar;107(2):279–283. doi: 10.1042/bj1070279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Griffin M. J., Cox R. P. Studies on the mechanism of hormonal induction of alkaline phosphatase in human cell cultures. I. Effects of puromycin and actinomycin D. J Cell Biol. 1966 Apr;29(1):1–9. doi: 10.1083/jcb.29.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Griffin M. J., Price G. H., Bazzell K. L. A study of adenosine 3':5'-cyclic monophosphate, sodium butyrate and cortisol as inducers of HeLa alkaline phosphatase. Arch Biochem Biophys. 1974 Oct;164(2):619–623. doi: 10.1016/0003-9861(74)90073-3. [DOI] [PubMed] [Google Scholar]
  17. Kelly S. H., Sperow J. W., Butler L. G. Stimulation of alkaline phosphatase by analogs of inorganic pyrophosphate. Biochemistry. 1974 Aug 13;13(17):3503–3505. doi: 10.1021/bi00714a014. [DOI] [PubMed] [Google Scholar]
  18. Koyama H., Kato R., Ono T. Induction of alkaline phosphatase by cyclic AMP or its dibutyryl derivative in a hybrid line between mouse and Chinese hamster in culture. Biochem Biophys Res Commun. 1972 Jan 14;46(1):305–311. doi: 10.1016/0006-291x(72)90663-8. [DOI] [PubMed] [Google Scholar]
  19. Koyama H., Ono T. Further studies on the induction of alkaline phosphatase by 5-bromodeoxyuridine in a hybrid line between mouse and Chinese hamster in culture. Biochim Biophys Acta. 1972 May 16;264(3):497–507. doi: 10.1016/0304-4165(72)90013-x. [DOI] [PubMed] [Google Scholar]
  20. Melnykovych G., Lopez I. C. Rapid cell surface-related stimulation of alkaline phosphatase in HeLa cells by dimethyl DL-2,3-distearoylotypropyl-2'-hydroxyethylammonium acetate (Rosenthal's inhibitor). J Cell Physiol. 1977 Jul;92(1):91–96. doi: 10.1002/jcp.1040920111. [DOI] [PubMed] [Google Scholar]
  21. Morgan D. B., Monod A., Russel R. G., Fleisch H. Influence of dichloromethylene diphosphonate (Cl2MDP) and calcitonin on bone resorption, lactate production and phosphatase and pyrophosphatase content of mouse calvaria treated with parathyroid hormone in vitro. Calcif Tissue Res. 1973 Dec 31;13(4):287–294. doi: 10.1007/BF02015418. [DOI] [PubMed] [Google Scholar]
  22. NITOWSKY H. M., HERZ F., GELLER S. INDUCTION OF ALKALINE PHOSPHATASE IN DISPERSED CELL CULTURES BY CHANGES IN OSMOLARITY. Biochem Biophys Res Commun. 1963 Aug 1;12:293–299. doi: 10.1016/0006-291x(63)90299-7. [DOI] [PubMed] [Google Scholar]
  23. Russell R. G., Fleisch H. Pyrophosphate and diphosphonates in skeletal metabolism. Physiological, clinical and therapeutic aspects. Clin Orthop Relat Res. 1975 May;(108):241–263. doi: 10.1097/00003086-197505000-00038. [DOI] [PubMed] [Google Scholar]
  24. Speeg K. V., Jr, Azizkhan J. C., Stromberg K. Characteristics of alkaline phosphatase from two continuous lines of human choriocarcinoma cells. Exp Cell Res. 1977 Mar 1;105(1):199–205. doi: 10.1016/0014-4827(77)90166-5. [DOI] [PubMed] [Google Scholar]
  25. Thomas M. L., Goz B. Parathyroid hormone inhibition of induction of alkaline phosphatase activity by hydrocortisone and 5-iodo-2'-deoxyuridine in HeLa cells. Exp Cell Res. 1976 Sep;101(2):422–424. doi: 10.1016/0014-4827(76)90396-7. [DOI] [PubMed] [Google Scholar]
  26. Wöltgens J. H., Bonting S. L., Bijvoet O. L. Inorganic pyrophosphatase in mineralizing hamster molars. 3. Influence of diphosphonates. Calcif Tissue Res. 1973;13(2):151–157. doi: 10.1007/BF02015405. [DOI] [PubMed] [Google Scholar]

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