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. 1991 Dec;88(6):1841–1847. doi: 10.1172/JCI115505

Abundant expression of parathyroid hormone-related protein in primary rat aortic smooth muscle cells accompanies serum-induced proliferation.

T Hongo 1, J Kupfer 1, H Enomoto 1, B Sharifi 1, D Giannella-Neto 1, J S Forrester 1, F R Singer 1, D Goltzman 1, G N Hendy 1, C Pirola 1, et al.
PMCID: PMC295751  PMID: 1752945

Abstract

Parathyroid hormone-related protein (PTHrP), which is responsible for producing hypercalcemia in patients with humoral hypercalcemia of malignancy, has recently been identified in several normal tissues. Because PTHrP, like parathyroid hormone (PTH), is known to exhibit vasodilatory properties, we investigated the expression and regulation of PTHrP mRNA in cultured rat aortic smooth muscle cells (SMC). We report here that PTHrP mRNA is expressed in SMC and is markedly induced by serum in a time- and concentration-dependent fashion. Addition of 10% fetal calf serum to serum-deprived, confluent cells, resulted in a marked induction of PTHrP mRNA by 2 h with a peak at 4-6 h. PTHrP was detected in SMC by immunocytochemistry and radioimmunoassay of conditioned medium, and was shown to be up-regulated within 24 h after the addition of serum. The serum induction of PTHrP mRNA was blocked by actinomycin D and by cycloheximide indicating the need for protein synthesis to evoke the serum effect on PTHrP gene transcription. In addition, treatment with dexamethasone, which has been previously shown to reduce the constitutive expression of PTHrP in human cancer cells, also blunted the serum induction of PTHrP mRNA in SMC. Treatment of quiescent cells with the serum mitogens platelet-derived growth factor or insulin-like growth factor-I had no effect on PTHrP, whereas the vasoactive peptides endothelin, norepinephrine and thrombin stimulated PTHrP expression. Exogenous addition of recombinant PTHrP-(1-141) had no significant effect on SMC DNA synthesis as measured by [3H]thymidine incorporation. In summary, the abundance of PTHrP mRNA and the characteristics of its regulation in SMC suggest a major role for PTHrP as a local modulator in vascular smooth muscle.

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Selected References

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

  1. Adachi N., Yamaguchi K., Miyake Y., Honda S., Nagasaki K., Akiyama Y., Adachi I., Abe K. Parathyroid hormone-related protein is a possible autocrine growth inhibitor for lymphocytes. Biochem Biophys Res Commun. 1990 Feb 14;166(3):1088–1094. doi: 10.1016/0006-291x(90)90978-v. [DOI] [PubMed] [Google Scholar]
  2. Cercek B., Fishbein M. C., Forrester J. S., Helfant R. H., Fagin J. A. Induction of insulin-like growth factor I messenger RNA in rat aorta after balloon denudation. Circ Res. 1990 Jun;66(6):1755–1760. doi: 10.1161/01.res.66.6.1755. [DOI] [PubMed] [Google Scholar]
  3. Chamley-Campbell J., Campbell G. R., Ross R. The smooth muscle cell in culture. Physiol Rev. 1979 Jan;59(1):1–61. doi: 10.1152/physrev.1979.59.1.1. [DOI] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Clemens T. L., Garrett K. P., Zhou X. Y., Pike J. W., Haussler M. R., Dempster D. W. Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. Endocrinology. 1988 Apr;122(4):1224–1230. doi: 10.1210/endo-122-4-1224. [DOI] [PubMed] [Google Scholar]
  6. Clemmons D. R., Van Wyk J. J. Evidence for a functional role of endogenously produced somatomedinlike peptides in the regulation of DNA synthesis in cultured human fibroblasts and porcine smooth muscle cells. J Clin Invest. 1985 Jun;75(6):1914–1918. doi: 10.1172/JCI111906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Danielson P. E., Forss-Petter S., Brow M. A., Calavetta L., Douglass J., Milner R. J., Sutcliffe J. G. p1B15: a cDNA clone of the rat mRNA encoding cyclophilin. DNA. 1988 May;7(4):261–267. doi: 10.1089/dna.1988.7.261. [DOI] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  9. Horiuchi N., Caulfield M. P., Fisher J. E., Goldman M. E., McKee R. L., Reagan J. E., Levy J. J., Nutt R. F., Rodan S. B., Schofield T. L. Similarity of synthetic peptide from human tumor to parathyroid hormone in vivo and in vitro. Science. 1987 Dec 11;238(4833):1566–1568. doi: 10.1126/science.3685994. [DOI] [PubMed] [Google Scholar]
  10. Ikeda K., Lu C., Weir E. C., Mangin M., Broadus A. E. Regulation of parathyroid hormone-related peptide gene expression by cycloheximide. J Biol Chem. 1990 Apr 5;265(10):5398–5402. [PubMed] [Google Scholar]
  11. Ikeda K., Lu C., Weir E. C., Mangin M., Broadus A. E. Transcriptional regulation of the parathyroid hormone-related peptide gene by glucocorticoids and vitamin D in a human C-cell line. J Biol Chem. 1989 Sep 25;264(27):15743–15746. [PubMed] [Google Scholar]
  12. Ikeda K., Weir E. C., Mangin M., Dannies P. S., Kinder B., Deftos L. J., Brown E. M., Broadus A. E. Expression of messenger ribonucleic acids encoding a parathyroid hormone-like peptide in normal human and animal tissues with abnormal expression in human parathyroid adenomas. Mol Endocrinol. 1988 Dec;2(12):1230–1236. doi: 10.1210/mend-2-12-1230. [DOI] [PubMed] [Google Scholar]
  13. Jones S. D., Hall D. J., Rollins B. J., Stiles C. D. Platelet-derived growth factor generates at least two distinct intracellular signals that modulate gene expression. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 1):531–536. doi: 10.1101/sqb.1988.053.01.061. [DOI] [PubMed] [Google Scholar]
  14. Karaplis A. C., Yasuda T., Hendy G. N., Goltzman D., Banville D. Gene-encoding parathyroid hormone-like peptide: nucleotide sequence of the rat gene and comparison with the human homologue. Mol Endocrinol. 1990 Mar;4(3):441–446. doi: 10.1210/mend-4-3-441. [DOI] [PubMed] [Google Scholar]
  15. Kemp B. E., Moseley J. M., Rodda C. P., Ebeling P. R., Wettenhall R. E., Stapleton D., Diefenbach-Jagger H., Ure F., Michelangeli V. P., Simmons H. A. Parathyroid hormone-related protein of malignancy: active synthetic fragments. Science. 1987 Dec 11;238(4833):1568–1570. doi: 10.1126/science.3685995. [DOI] [PubMed] [Google Scholar]
  16. Kindy M. S., Sonenshein G. E. Regulation of oncogene expression in cultured aortic smooth muscle cells. Post-transcriptional control of c-myc mRNA. J Biol Chem. 1986 Sep 25;261(27):12865–12868. [PubMed] [Google Scholar]
  17. Lu C., Ikeda K., Deftos L. J., Gazdar A. F., Mangin M., Broadus A. E. Glucocorticoid regulation of parathyroid hormone-related peptide gene transcription in a human neuroendocrine cell line. Mol Endocrinol. 1989 Dec;3(12):2034–2040. doi: 10.1210/mend-3-12-2034. [DOI] [PubMed] [Google Scholar]
  18. Mangin M., Ikeda K., Dreyer B. E., Broadus A. E. Isolation and characterization of the human parathyroid hormone-like peptide gene. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2408–2412. doi: 10.1073/pnas.86.7.2408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mok L. L., Nickols G. A., Thompson J. C., Cooper C. W. Parathyroid hormone as a smooth muscle relaxant. Endocr Rev. 1989 Nov;10(4):420–436. doi: 10.1210/edrv-10-4-420. [DOI] [PubMed] [Google Scholar]
  20. Moseley J. M., Kubota M., Diefenbach-Jagger H., Wettenhall R. E., Kemp B. E., Suva L. J., Rodda C. P., Ebeling P. R., Hudson P. J., Zajac J. D. Parathyroid hormone-related protein purified from a human lung cancer cell line. Proc Natl Acad Sci U S A. 1987 Jul;84(14):5048–5052. doi: 10.1073/pnas.84.14.5048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reed J. C., Alpers J. D., Nowell P. C., Hoover R. G. Sequential expression of protooncogenes during lectin-stimulated mitogenesis of normal human lymphocytes. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3982–3986. doi: 10.1073/pnas.83.11.3982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rice B. F., Ponthier R. L., Jr, Miller M. C., 3rd Hypercalcemia and neoplasia: a model system. Endocrinology. 1971 May;88(5):1210–1216. doi: 10.1210/endo-88-5-1210. [DOI] [PubMed] [Google Scholar]
  23. Rodda C. P., Kubota M., Heath J. A., Ebeling P. R., Moseley J. M., Care A. D., Caple I. W., Martin T. J. Evidence for a novel parathyroid hormone-related protein in fetal lamb parathyroid glands and sheep placenta: comparisons with a similar protein implicated in humoral hypercalcaemia of malignancy. J Endocrinol. 1988 May;117(2):261–271. doi: 10.1677/joe.0.1170261. [DOI] [PubMed] [Google Scholar]
  24. Stewart A. F., Wu T., Goumas D., Burtis W. J., Broadus A. E. N-terminal amino acid sequence of two novel tumor-derived adenylate cyclase-stimulating proteins: identification of parathyroid hormone-like and parathyroid hormone-unlike domains. Biochem Biophys Res Commun. 1987 Jul 31;146(2):672–678. doi: 10.1016/0006-291x(87)90581-x. [DOI] [PubMed] [Google Scholar]
  25. Strewler G. J., Stern P. H., Jacobs J. W., Eveloff J., Klein R. F., Leung S. C., Rosenblatt M., Nissenson R. A. Parathyroid hormonelike protein from human renal carcinoma cells. Structural and functional homology with parathyroid hormone. J Clin Invest. 1987 Dec;80(6):1803–1807. doi: 10.1172/JCI113275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Thiede M. A., Daifotis A. G., Weir E. C., Brines M. L., Burtis W. J., Ikeda K., Dreyer B. E., Garfield R. E., Broadus A. E. Intrauterine occupancy controls expression of the parathyroid hormone-related peptide gene in preterm rat myometrium. Proc Natl Acad Sci U S A. 1990 Sep;87(18):6969–6973. doi: 10.1073/pnas.87.18.6969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Thiede M. A., Rodan G. A. Expression of a calcium-mobilizing parathyroid hormone-like peptide in lactating mammary tissue. Science. 1988 Oct 14;242(4876):278–280. doi: 10.1126/science.3175653. [DOI] [PubMed] [Google Scholar]
  28. Walker L. N., Bowen-Pope D. F., Ross R., Reidy M. A. Production of platelet-derived growth factor-like molecules by cultured arterial smooth muscle cells accompanies proliferation after arterial injury. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7311–7315. doi: 10.1073/pnas.83.19.7311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yasuda T., Banville D., Hendy G. N., Goltzman D. Characterization of the human parathyroid hormone-like peptide gene. Functional and evolutionary aspects. J Biol Chem. 1989 May 5;264(13):7720–7725. [PubMed] [Google Scholar]
  30. Yasuda T., Banville D., Rabbani S. A., Hendy G. N., Goltzman D. Rat parathyroid hormone-like peptide: comparison with the human homologue and expression in malignant and normal tissue. Mol Endocrinol. 1989 Mar;3(3):518–525. doi: 10.1210/mend-3-3-518. [DOI] [PubMed] [Google Scholar]
  31. Yates A. J., Gutierrez G. E., Smolens P., Travis P. S., Katz M. S., Aufdemorte T. B., Boyce B. F., Hymer T. K., Poser J. W., Mundy G. R. Effects of a synthetic peptide of a parathyroid hormone-related protein on calcium homeostasis, renal tubular calcium reabsorption, and bone metabolism in vivo and in vitro in rodents. J Clin Invest. 1988 Mar;81(3):932–938. doi: 10.1172/JCI113406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zhou X. Y., Dempster D. W., Marion S. L., Pike J. W., Haussler M. R., Clemens T. L. Bone vitamin D-dependent calcium-binding protein is localized in chondrocytes of growth-plate cartilage. Calcif Tissue Int. 1986 Apr;38(4):244–247. doi: 10.1007/BF02556718. [DOI] [PubMed] [Google Scholar]

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