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Effects of hydroxyurea and cyclic adenosine monophosphate/protein kinase A inhibitors on the expression of the human chorionic gonadotropin alpha subunit and c-myc genes in choriocarcinoma

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Abstract

We have previously shown that treatment of choriocarcinoma cells with methotrexate or hydroxyurea leads to both cessation of cell growth, accompanied by repression of c-myc oncogene expression, and induction of genes associated with the placental phenotype, including both subunits of human chorionic gonadotropin (hCG) and placental alkaline phosphatase. Since the genes induced by these antimetabolites are also cyclic AMP inducible, we hypothesized that these antimetabolites may cause activation of the cyclic AMP/protein kinase A pathway, suppressing genes associated with cellular proliferation and inducing placental gene expression. Three inhibitors of the cyclic AMP/protein kinase A pathway were assayed for their ability to inhibit the induction of the human chorionic gonadotropin alpha gene by hydroxyurea, and none of these inhibitors eliminated this induction. In addition, blockade of the cyclic AMP/protein kinase A pathway did not reverse the suppression of c-myc by hydroxyurea. The results of the inhibitor studies suggest that hydroxyurea acts independently of the protein kinase A pathway to stimulate gene expression, and that suppression of c-myc is insufficient to cause the induction of the human chorionic gonadotropin alpha gene by hydroxyurea.

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References

  1. Ringler G.E., Strauss J.F. III. In vitro systems for the study of human placental endocrine function. Endocrine Rev. 11: 105, 1990.

    Article  CAS  Google Scholar 

  2. Goldstein D., Berkowitz R.S. Gestational Trophoblastic Neoplasms: Clinical principles of diagnosis and management. W.B. Saunders Company, Philadelphia 1982.

    Google Scholar 

  3. Li M.C., Hertz R., Spencer D.B. Effect of methotrexate therapy upon choriocarcinoma and chorioadenoma. Proc. Soc. Exp. Biol. Med. 93: 361, 1956.

    Article  PubMed  Google Scholar 

  4. Berkowitz R.S., Goldstein D.P., Bernstein M.R. Ten years experience with methotrexate and folinic acid as primary therapy for gestational trophoblastic disease. Gynecol. Oncol. 23: 111, 1986.

    Article  PubMed  CAS  Google Scholar 

  5. Arbiser J.L., Arbiser Z.K., Majzoub J.A. Regulation of gene expression in choriocarcinoma by methotrexate and hydroxyurea. Endocrinol. 128: 972, 1991

    Article  CAS  Google Scholar 

  6. Taylor R.N., Newman E., Chen S. Forskolin and methotrexate induce an intermediate trophoblast phenotype in cultured human choriocarcinoma cells. Am J. Obstet. Gynecol. 164: 204, 1991.

    Article  PubMed  CAS  Google Scholar 

  7. Speeg K. Jr., Azizkhan J., Stromberg K. The stimulation by methotrexate of human chorionic gonadotropin and placental alkaline phosphatase in cultured choriocarcinoma cells. Cancer Res. 36: 4570, 1976

    PubMed  CAS  Google Scholar 

  8. Azizkhan J.,Speeg K. Jr., Stromberg K., Goode D. Stimulation of human chorionic gonadotropin by JAr line choriocarcinoma after inhibition of DNA synthesis. Cancer Res. 39: 1952, 1979.

    PubMed  CAS  Google Scholar 

  9. Chou J.Y., Robinson J.C. Induction of alkaline phosphatase in choriocarcinoma cells by 1-barabinofuranosylcytosine, mitomycin C,phleomycin, and cyclic nucleotides. J-Cell. Physiol. 92: 221, 1977

    Article  PubMed  CAS  Google Scholar 

  10. Friedman S., Skehan P. Morphological differentiation of human choriocarcinoma cells induced by methotrexate. Cancer Res. 39: 1960, 1979

    PubMed  CAS  Google Scholar 

  11. Browne P., Bagshawe K. D. Enhancement of human chorionic gonadotropin production by antimetabolites. Br. J. Cancer 46: 22, 1982

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  12. Burres N., Cass C. Inhibition of methotrexate-induced differentiation of cultured human choriocarcinoma cells by thymidine. Cancer Res. 47: 5059, 1987.

    PubMed  CAS  Google Scholar 

  13. Jameson J.L., Jaffe R., Gleason S., Habener J.F. Transcriptional regulation of chorionic gonadotropin alpha and beta subunit gene expression by 8-bro-mo-adenosine 3′5′-monophosphate. Endocrinology 119: 2560, 1986.

    Article  PubMed  CAS  Google Scholar 

  14. Silver B.J., Bokar J.A., Virgin J.B., Vallen E.A., Milsted A., Nilson J.H. Cyclic AMP regulation of the human glycoprotein alpha subunit gene is mediated by an 18-base pair element. Proc. Natl. Acad. Sci. USA 84: 2198, 1987.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  15. Hamilton T.A., Tin. A.W., Sussman H.H. Regulation of alkaline phosphatase expression in human choriocarcinoma cell lines. Proc. Natl. Acad.Sci. USA: 323, 1979.

  16. Albert DA, Gudas L.J., Nodzenski E. Deoxyribonucleotide metabolism and cyclic AMP resistance in hydroxyurea resistant S49 T-lymphoma cells. J. Cell. Physiol. 130: 262, 1987.

    Article  PubMed  CAS  Google Scholar 

  17. Krakoff I. H., Brown N. C., Reichard P. Inhibition of ribonucleoside diphosphate reductase by hydroxyurea. Cancer Res. 28: 1559, 1968.

    PubMed  CAS  Google Scholar 

  18. Cole M.D. The myc oncogene: Its role in transformation and differentiation. Ann. Rev. Genet. 20: 361,1986

    Article  PubMed  CAS  Google Scholar 

  19. Pfeifer-Ohlsson S., Goustin A. S., Rydnert J., Wahlstrom T., Bjersing L., Stehelin D., Ohlsson R. Spatial and temporal pattern of cellular myc oncogene expression in developing human placenta: Implications for embryonic cell proliferation. Cell 38: 585, 1984.

    Article  PubMed  CAS  Google Scholar 

  20. Sarkar S., Kacinski B.M., Kohorn E.I., Merino M.J., Carter D., Blakemore K.J. Demonstration of myc and ras oncogene expression by hybridization in situ in hydatidiform mole and in the BeWo choriocarcinoma cell line. Am. J. Obstet. Gynecol. 154: 390, 1986.

    Article  PubMed  CAS  Google Scholar 

  21. Goldberg H., Clayman P., Skorecki K. Mechanism of Li inhibition of vasopressin sensitive adenylate cyclase in cultured renal epithelial cells. Am. J. Physioll 255: F995, 1988.

    CAS  Google Scholar 

  22. Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry 23: 5036, 1984.

    Article  PubMed  CAS  Google Scholar 

  23. Parker-Botelho L.H., Rothermel J.D., Coombs R., Jastorff B. cAMP analog antagonists of cAMP action. Methods Enzymol. 159: 159, 1988

    Article  Google Scholar 

  24. Kohler P., Bridson W. Isolation of hormone-producing clonal lines of human choriocarcinoma. J. Clin. Endocrinol. Metab. 32: 683, 1971.

    Article  PubMed  CAS  Google Scholar 

  25. Chirgwin J.M., Przybla A.E., MacDonald R.J., Rutter W.J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294, 1979.

    Article  PubMed  CAS  Google Scholar 

  26. Fiddes J.C., Goodman H.M. Isolation, cloning, and sequence analysis of the cDNA for the alpha subunit of human chorionic gonadotropin. Nature 281: 351, 1979.

    Article  PubMed  CAS  Google Scholar 

  27. Adler G.K., Smas C.M., Majzoub J.A. Expression and dexamethasone regulation of the human corticotropinreleasing hormone gene in a mouse anterior pituitary cell line. J. Biol. Chem. 263: 5846, 1988.

    PubMed  CAS  Google Scholar 

  28. Fiddes J.C., Seeburg P.H., DeNoto F.M., Hallewell R., Baxter J.D., Goodman H.M. Structure of genes for human growth hormone and chorionic somatomammotropin. Proc. Natl. Acad. Sci. USA 76: 4294, 1979.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  29. Feinberg A., Vogelstein B. A technique for labelling DNA restriction fragments to high specific activity. Analyt. Biochem. 137: 266, 1984.

    Article  PubMed  CAS  Google Scholar 

  30. Melton D.A., Krieg P.A., Rebagliati M.R., Maniatis T., Zinn K., Green M.R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 12: 7035, 1984.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  31. Fenstermaker R. A., Milsted A., Virgin J., Miller W., Nilson J. H. The transcriptional response of the human chorionic gonadotropin beta-subunit gene to cAMP is cycloheximide sensitive and is mediated by cis-acting sequences different from that found in the alpha-subunit gene. Mol. Endocrinol. 3: 1070, 1989

    Article  PubMed  CAS  Google Scholar 

  32. Correll L., Woodford T., Corbin J., Mellon P., McKnight G.S. Functional characterization of cAMP-binding mutations in type I protein kinase. J. Biol. Chem. 264: 16672, 1989.

    PubMed  CAS  Google Scholar 

  33. Berridge M.J. Inositol trisphosphate and diacylglycerol: Two interacting second messengers. Ann. Rev. Biochem. 56: 159, 1987.

    Article  PubMed  CAS  Google Scholar 

  34. Pereira M.E., Segaloff D.L., Ascoli M., Eckstein F. Inhibition of choriogonadotrpin-activated steroidogenesis in cultured leydig tumor cells by the Rp diastereoisomer of adenosine 3′5′-cyclic phosphorothioate. J. Biol. Chem. 262: 6093, 1987.

    PubMed  CAS  Google Scholar 

  35. Grove J.R., Deutsch P.J., Price D.J., Habener J.F. Avruch Plasmids encoding PKI(1–31), a specific inhibitor of cAMP-stimulated gene expression, inhibit the basal transcriptional activity of some but not all cAMP-regulated DNA response elements in JEG-3 cells. J. Biol. Chem. 264: 19506, 1989.

    PubMed  CAS  Google Scholar 

  36. Moore J.J., Lundgren D.W., Moore R.M., Andersen B. Polyamines control human chorionic gonadotropin production in the JEG-3 choriocarcinoma cell. J. Biol. Chem. 263: 12765, 1988.

    PubMed  CAS  Google Scholar 

  37. Celano P., Baylin S.B., Giardiello F.M., Nelkin B.D., Casero R. Effect of polyamine depletion on c-myc expression in human colon carcinoma cells. J. Biol. Chem. 263: 5491, 1988.

    PubMed  CAS  Google Scholar 

  38. Bodner A., Ting R., Gallo R. Induction of differentiation of human promyelocytic leukemia cells (HL-60) by nucleosides and methotrexate. J. Natl. Cancer Inst. 67: 1025, 1981.

    PubMed  CAS  Google Scholar 

  39. Sokoloski J.A., Beardsley G.P., Sartorelli A.C. Mechanism of the induction of the differentiation of HL-60 leukemia cells by antifolates. Cancer. Commun. 1: 199, 1989.

    PubMed  CAS  Google Scholar 

  40. Galanello R., Stamatoyannopoulos G., Papayannopoulou T. Mechanism of HbF stimulation by S-stage compounds: in vitro studies with bone marrow cells exposed to 5-azacytidine, Ara-C, or hydroxyurea. J.Clin. Invest. 81: 1209, 1988.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

  41. Hussa R.O., Pattillo RA, Ruckert A.C., Scheuerman K.W. Effects of butyrate and dibutyryl cyclic AMP on hCG secreting trophoblastic and non-trophoblastic cells. J. Clin. Endocrinol. Metab. 46: 69, 1978.

    Article  PubMed  CAS  Google Scholar 

  42. Ghosh N.K., Rukenstein A., Cox R.P. Induction of human choriogonadotropin in HeLa cell cultures by alpiphatic monocarboxylates and inhibitors of deoxyribonucleic acid synthesis. Biochem. J. 166: 265, 1977.

    PubMed Central  PubMed  CAS  Google Scholar 

  43. Darnell R. B., Boime I. Differential expression of the human gonadotropin alpha gene in ectopic and eutopic cells. Mol. Cell. Biol. 5: 3157, 1985.

    PubMed Central  PubMed  CAS  Google Scholar 

  44. Boime I., Landefeld T., NcQueen S., McWilliams D. The biosynthesis of chorionic gonadotropin and placental lactogen in first and third trimester human placenta. In: McKerns K. (Ed), Structure and function of the gonadotropins. Plenum Press, New York, 1978 p. 235.

    Chapter  Google Scholar 

  45. Frim D.F., Emanuel R.L., Robinson B., Adler G., Majzoub J.A. Characterization and gestational regulation of corticotropinreleasing hormone messenger RNA in human placenta. J.Clin. Invest. 82: 287, 1988.

    Article  PubMed Central  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Division of Endocrinology, Children Hospital, 300 Mongwood Ave, Boston, MA, 02115, USA

    J. L. Arbiser, Z. K. Arbiser & J. A. Majzoub

  2. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    J. L. Arbiser

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  1. J. L. Arbiser
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  2. Z. K. Arbiser
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  3. J. A. Majzoub
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Additional information

This work was supported in part by Medical Scientist Training Program Grant 5T-32-GH-07753-09 (to JLA) and Grant RO1-HD-24704 (to JM) from the NIH.

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Arbiser, J.L., Arbiser, Z.K. & Majzoub, J.A. Effects of hydroxyurea and cyclic adenosine monophosphate/protein kinase A inhibitors on the expression of the human chorionic gonadotropin alpha subunit and c-myc genes in choriocarcinoma. J Endocrinol Invest 16, 849–856 (1993). https://doi.org/10.1007/BF03348942

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