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. 1981 Jul 15;198(1):29–35. doi: 10.1042/bj1980029

The effect of bromodeoxyuridine and dibutyryl cyclic AMP on the induction of placental alkaline phosphate in human choriocarcinoma cells.

T A Hamilton, H H Sussman
PMCID: PMC1163207  PMID: 6275841

Abstract

The effect of 5-bromo-2'-deoxyuridine (BrdUrd) and dibutyryl cyclic AMP (Bt2cAMP) on the expression of the placental isoenzyme of human alkaline phosphatase was examined in BeWo choriocarcinoma cells. By using a combination of specific immunoprecipitation and polyacrylamide-gel electrophoresis of cells labelled either metabolically with [35S]methionine or cell-surface-labelled with 125I, both BrdUrd (5 micrograms/ml) and 1 mM-Bt2cAMP were shown to result in the enhanced accumulation of a specific protein. This protein has immunochemical identity and co-electrophoreses with placental alkaline phosphatase in two-dimensional gels. These results clearly demonstrate that the induction of placental alkaline phosphatase activity in choriocarcinoma cells treated with these agents is a consequence of the accumulation of specific enzyme protein rather than of altered catalytic activity.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Badger K. S., Sussman H. H. Structural evidence that human liver and placental alkaline phosphatase isoenzymes are coded by different genes. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2201–2205. doi: 10.1073/pnas.73.7.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Fishman W. H., Inglis N. I., Stolbach L. L., Krant M. J. A serum alkaline phosphatase isoenzyme of human neoplastic cell origin. Cancer Res. 1968 Jan;28(1):150–154. [PubMed] [Google Scholar]
  3. Ghosh N. K., Rukenstein A., Baltimore R., Cox R. P. Studies on hormonal induction of alkaline phosphatase in HeLa cell cultures. Kinetic, thermodynamic and electrophoretic properties of induced and base-level enzymes. Biochim Biophys Acta. 1972 Nov 24;286(1):175–185. doi: 10.1016/0304-4165(72)90103-1. [DOI] [PubMed] [Google Scholar]
  4. Goz B., Walker K. P. An immunological and biochemical analysis of alkaline phosphatase in HeLa cells exposed to 5-iodo-2'-deoxyuridine. Biochem Pharmacol. 1978 Feb 15;27(4):431–436. doi: 10.1016/0006-2952(78)90372-6. [DOI] [PubMed] [Google Scholar]
  5. Greene P. J., Sussman H. H. Structual comparison of ectopic and normal placental alkaline phosphatase. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2936–2940. doi: 10.1073/pnas.70.10.2936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hamilton T. A., Tin A. W., Sussman H. H. Regulation of alkaline phosphatase expression in human choriocarcinoma cell lines. Proc Natl Acad Sci U S A. 1979 Jan;76(1):323–327. doi: 10.1073/pnas.76.1.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hamilton T. A., Wada H. G., Sussman H. H. Human placental cell surface antigens:expression by cultured cells of diverse phenotypic origin. J Supramol Struct. 1979;11(4):503–515. doi: 10.1002/jss.400110409. [DOI] [PubMed] [Google Scholar]
  8. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  11. McKenna M. J., Hamilton T. A., Sussman H. H. Comparison of human alkaline phosphatase isoenzymes. Structural evidence for three protein classes. Biochem J. 1979 Jul 1;181(1):67–73. doi: 10.1042/bj1810067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Morrow J., Stocco D. M., Fralick J. A. The requirement of DNA synthesis for the induction of alkaline phosphatase by bromodeoxyuridine in a derivative of the HeLa cell line. J Cell Physiol. 1979 Feb;98(2):427–436. doi: 10.1002/jcp.1040980218. [DOI] [PubMed] [Google Scholar]
  13. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  14. Singer R. M., Herz F. Multiple effects of glucocorticoids and hyperosmolality on isoenzyme expression in cultured KB cells. J Natl Cancer Inst. 1978 Mar;60(3):573–577. doi: 10.1093/jnci/60.3.573. [DOI] [PubMed] [Google Scholar]
  15. Smith C. H., Nelson D. M., King B. F., Donohue T. M., Ruzycki S., Kelley L. K. Characterization of a microvillous membrane preparation from human placental syncytiotrophoblast: a morphologic, biochemical, and physiologic, study. Am J Obstet Gynecol. 1977 May 15;128(2):190–196. doi: 10.1016/0002-9378(77)90686-x. [DOI] [PubMed] [Google Scholar]
  16. Sussman H. H., Small P. A., Jr, Cotlove E. Human alkaline phosphatase. Immunochemical identification of organ-specific isoenzymes. J Biol Chem. 1968 Jan 10;243(1):160–166. [PubMed] [Google Scholar]
  17. Wada H. G., Górnicki Z., Sussman H. H. The sialoglycoprotein subunits of human placental brush border membranes characterized by two-two-dimensional electrophoresis. J Supramol Struct. 1977;6(4):473–484. doi: 10.1002/jss.400060402. [DOI] [PubMed] [Google Scholar]
  18. Wada H. G., Hass P. E., Sussman H. H. Characterization of antigenic sialoglycoprotein subunits of the placental brush border membranes: comparison with liver and kidney membrane subunits by two-dimensional electrophoresis. J Supramol Struct. 1979;10(3):287–305. doi: 10.1002/jss.400100303. [DOI] [PubMed] [Google Scholar]
  19. Wada H. G., Shindelman J. E., Ortmeyer A. E., Sussman H. H. Demonstration of placental alkaline phosphatase in human breast cancer. Int J Cancer. 1979 Jun 15;23(6):781–787. doi: 10.1002/ijc.2910230608. [DOI] [PubMed] [Google Scholar]

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