Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Mar 15;97(6):1429–1435. doi: 10.1172/JCI118564

Inhibition of TGF-alpha gene expression by vitamin A in airway epithelium.

L A Miller 1, Y H Zhao 1, R Wu 1
PMCID: PMC507202  PMID: 8617875

Abstract

The autocrine/paracrine growth mechanism has been implicated in the regulation of bronchial epithelial cell proliferation. By inhibiting the expression of the transforming growth factor-alpha (TGF-alpha) gene product, vitamin A is able to suppress the proliferation of tracheobronchial epithelial cells in culture. Similar repressions in TGF-alpha mRNA levels by retinol were observed in airway explant cultures and in a cell line immortalized from normal human bronchial epithelial cells. Both the nuclear run-on transcriptional assay and the transfection study with the chimeric construct of the TGF-alpha promoter and chloramphenicol acetyltransferase reporter gene partly suggest a transcriptional downregulation mechanism of TGF-alpha gene expression by the retinol treatment; however, this inhibition at the transcriptional level cannot account for the total inhibition at the mRNA level. These results suggest that a downregulation of the expression of the TGF-alpha gene at the transcriptional and post-transcriptional levels by vitamin A may precede the essential event associated with the homeostasis of normal conducting airway epithelium.

Full Text

The Full Text of this article is available as a PDF (335.6 KB).

Selected References

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

  1. An G., Huang T. H., Tesfaigzi J., Garcia-Heras J., Ledbetter D. H., Carlson D. M., Wu R. An unusual expression of a squamous cell marker, small proline-rich protein gene, in tracheobronchial epithelium: differential regulation and gene mapping. Am J Respir Cell Mol Biol. 1992 Jul;7(1):104–111. doi: 10.1165/ajrcmb/7.1.104. [DOI] [PubMed] [Google Scholar]
  2. An G., Tesfaigzi J., Chuu Y. J., Wu R. Isolation and characterization of the human spr1 gene and its regulation of expression by phorbol ester and cyclic AMP. J Biol Chem. 1993 May 25;268(15):10977–10982. [PubMed] [Google Scholar]
  3. An G., Wu R. Thioredoxin gene expression is transcriptionally up-regulated by retinol in monkey conducting airway epithelial cells. Biochem Biophys Res Commun. 1992 Feb 28;183(1):170–175. doi: 10.1016/0006-291x(92)91624-y. [DOI] [PubMed] [Google Scholar]
  4. Brand N., Petkovich M., Krust A., Chambon P., de Thé H., Marchio A., Tiollais P., Dejean A. Identification of a second human retinoic acid receptor. Nature. 1988 Apr 28;332(6167):850–853. doi: 10.1038/332850a0. [DOI] [PubMed] [Google Scholar]
  5. Brand N., Petkovich M., Krust A., Chambon P., de Thé H., Marchio A., Tiollais P., Dejean A. Identification of a second human retinoic acid receptor. Nature. 1988 Apr 28;332(6167):850–853. doi: 10.1038/332850a0. [DOI] [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Chopra D. P. Squamous metaplasia in organ cultures of vitamin A-deficient hamster trachea: cytokinetic and ultrastructural alterations. J Natl Cancer Inst. 1982 Oct;69(4):895–905. [PubMed] [Google Scholar]
  8. Derynck R., Roberts A. B., Winkler M. E., Chen E. Y., Goeddel D. V. Human transforming growth factor-alpha: precursor structure and expression in E. coli. Cell. 1984 Aug;38(1):287–297. doi: 10.1016/0092-8674(84)90550-6. [DOI] [PubMed] [Google Scholar]
  9. Elder J. T., Fisher G. J., Lindquist P. B., Bennett G. L., Pittelkow M. R., Coffey R. J., Jr, Ellingsworth L., Derynck R., Voorhees J. J. Overexpression of transforming growth factor alpha in psoriatic epidermis. Science. 1989 Feb 10;243(4892):811–814. doi: 10.1126/science.2916128. [DOI] [PubMed] [Google Scholar]
  10. Giguère V., Shago M., Zirngibl R., Tate P., Rossant J., Varmuza S. Identification of a novel isoform of the retinoic acid receptor gamma expressed in the mouse embryo. Mol Cell Biol. 1990 May;10(5):2335–2340. doi: 10.1128/mcb.10.5.2335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gilliland G., Perrin S., Blanchard K., Bunn H. F. Analysis of cytokine mRNA and DNA: detection and quantitation by competitive polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2725–2729. doi: 10.1073/pnas.87.7.2725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harris C. C., Sporn M. B., Kaufman D. G., Smith J. M., Jackson F. E., Saffiotti U. Histogenesis of squamous metaplasia in the hamster tracheal epithelium caused by vitamin A deficiency or benzo[a]pyrene-Ferric oxide. J Natl Cancer Inst. 1972 Mar;48(3):743–761. [PubMed] [Google Scholar]
  13. Huang T. H., Ann D. K., Zhang Y. J., Chang A. T., Crabb J. W., Wu R. Control of keratin gene expression by vitamin A in tracheobronchial epithelial cells. Am J Respir Cell Mol Biol. 1994 Feb;10(2):192–201. doi: 10.1165/ajrcmb.10.2.7509163. [DOI] [PubMed] [Google Scholar]
  14. Jakobovits E. B., Schlokat U., Vannice J. L., Derynck R., Levinson A. D. The human transforming growth factor alpha promoter directs transcription initiation from a single site in the absence of a TATA sequence. Mol Cell Biol. 1988 Dec;8(12):5549–5554. doi: 10.1128/mcb.8.12.5549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jetten A. M. Multistep process of squamous differentiation in tracheobronchial epithelial cells in vitro: analogy with epidermal differentiation. Environ Health Perspect. 1989 Mar;80:149–160. doi: 10.1289/ehp.8980149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kark J. D., Smith A. H., Switzer B. R., Hames C. G. Serum vitamin A (retinol) and cancer incidence in Evans County, Georgia. J Natl Cancer Inst. 1981 Jan;66(1):7–16. [PubMed] [Google Scholar]
  17. Keenan K. P., Combs J. W., McDowell E. M. Regeneration of hamster tracheal epithelium after mechanical injury. II. Multifocal lesions: stathmokinetic and autoradiographic studies of cell proliferation. Virchows Arch B Cell Pathol Incl Mol Pathol. 1982;41(3):215–229. doi: 10.1007/BF02890282. [DOI] [PubMed] [Google Scholar]
  18. Korc M., Chandrasekar B., Shah G. N. Differential binding and biological activities of epidermal growth factor and transforming growth factor alpha in a human pancreatic cancer cell line. Cancer Res. 1991 Dec 1;51(23 Pt 1):6243–6249. [PubMed] [Google Scholar]
  19. Krust A., Kastner P., Petkovich M., Zelent A., Chambon P. A third human retinoic acid receptor, hRAR-gamma. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5310–5314. doi: 10.1073/pnas.86.14.5310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kubiak J., Mitra M. M., Steve A. R., Hunt J. D., Davies P., Pitt B. R. Transforming growth factor-alpha gene expression in late-gestation fetal rat lung. Pediatr Res. 1992 Mar;31(3):286–290. doi: 10.1203/00006450-199203000-00019. [DOI] [PubMed] [Google Scholar]
  21. Mader S., Leroy P., Chen J. Y., Chambon P. Multiple parameters control the selectivity of nuclear receptors for their response elements. Selectivity and promiscuity in response element recognition by retinoic acid receptors and retinoid X receptors. J Biol Chem. 1993 Jan 5;268(1):591–600. [PubMed] [Google Scholar]
  22. Mangelsdorf D. J., Borgmeyer U., Heyman R. A., Zhou J. Y., Ong E. S., Oro A. E., Kakizuka A., Evans R. M. Characterization of three RXR genes that mediate the action of 9-cis retinoic acid. Genes Dev. 1992 Mar;6(3):329–344. doi: 10.1101/gad.6.3.329. [DOI] [PubMed] [Google Scholar]
  23. Mangelsdorf D. J., Ong E. S., Dyck J. A., Evans R. M. Nuclear receptor that identifies a novel retinoic acid response pathway. Nature. 1990 May 17;345(6272):224–229. doi: 10.1038/345224a0. [DOI] [PubMed] [Google Scholar]
  24. Mangelsdorf D. J. Vitamin A receptors. Nutr Rev. 1994 Feb;52(2 Pt 2):S32–S44. doi: 10.1111/j.1753-4887.1994.tb01385.x. [DOI] [PubMed] [Google Scholar]
  25. Marchok A. C., Cone V., Nettesheim P. Induction of squamous metaplasia (vitamin A deficiency) and hypersecretory activity in tracheal organ cultures. Lab Invest. 1975 Oct;33(4):451–460. [PubMed] [Google Scholar]
  26. Massagué J. Transforming growth factor-alpha. A model for membrane-anchored growth factors. J Biol Chem. 1990 Dec 15;265(35):21393–21396. [PubMed] [Google Scholar]
  27. Miller L. A., Cheng L. Z., Wu R. Inhibition of epidermal growth factor-like growth factor secretion in tracheobronchial epithelial cells by vitamin A. Cancer Res. 1993 Jun 1;53(11):2527–2533. [PubMed] [Google Scholar]
  28. Parham D. M., Jankowski J. Transforming growth factor alpha in epithelial proliferative diseases of the breast. J Clin Pathol. 1992 Jun;45(6):513–516. doi: 10.1136/jcp.45.6.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Petkovich M., Brand N. J., Krust A., Chambon P. A human retinoic acid receptor which belongs to the family of nuclear receptors. Nature. 1987 Dec 3;330(6147):444–450. doi: 10.1038/330444a0. [DOI] [PubMed] [Google Scholar]
  30. Petkovich M. Regulation of gene expression by vitamin A: the role of nuclear retinoic acid receptors. Annu Rev Nutr. 1992;12:443–471. doi: 10.1146/annurev.nu.12.070192.002303. [DOI] [PubMed] [Google Scholar]
  31. Pittelkow M. R., Lindquist P. B., Abraham R. T., Graves-Deal R., Derynck R., Coffey R. J., Jr Induction of transforming growth factor-alpha expression in human keratinocytes by phorbol esters. J Biol Chem. 1989 Mar 25;264(9):5164–5171. [PubMed] [Google Scholar]
  32. Raaberg L., Nexø E., Buckley S., Luo W., Snead M. L., Warburton D. Epidermal growth factor transcription, translation, and signal transduction by rat type II pneumocytes in culture. Am J Respir Cell Mol Biol. 1992 Jan;6(1):44–49. doi: 10.1165/ajrcmb/6.1.44. [DOI] [PubMed] [Google Scholar]
  33. Riedy M. C., Timm E. A., Jr, Stewart C. C. Quantitative RT-PCR for measuring gene expression. Biotechniques. 1995 Jan;18(1):70-4, 76. [PubMed] [Google Scholar]
  34. Sadiq H. F., Devaskar U. P. Glucocorticoids increase pulmonary epidermal growth factor receptors in female and male fetal rabbit. Biochem Biophys Res Commun. 1984 Feb 29;119(1):408–414. doi: 10.1016/0006-291x(84)91667-x. [DOI] [PubMed] [Google Scholar]
  35. Saffiotti U., Montesano R., Sellakumar A. R., Borg S. A. Experimental cancer of the lung. Inhibition by vitamin A of the induction of tracheobronchial squamous metaplasia and squamous cell tumors. Cancer. 1967 May;20(5):857–864. doi: 10.1002/1097-0142(1967)20:5<857::aid-cncr2820200545>3.0.co;2-3. [DOI] [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schüle R., Rangarajan P., Yang N., Kliewer S., Ransone L. J., Bolado J., Verma I. M., Evans R. M. Retinoic acid is a negative regulator of AP-1-responsive genes. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6092–6096. doi: 10.1073/pnas.88.14.6092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Seed B., Sheen J. Y. A simple phase-extraction assay for chloramphenicol acyltransferase activity. Gene. 1988 Jul 30;67(2):271–277. doi: 10.1016/0378-1119(88)90403-9. [DOI] [PubMed] [Google Scholar]
  39. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  40. Stahlman M. T., Orth D. N., Gray M. E. Immunocytochemical localization of epidermal growth factor in the developing human respiratory system and in acute and chronic lung disease in the neonate. Lab Invest. 1989 Apr;60(4):539–547. [PubMed] [Google Scholar]
  41. Strandjord T. P., Clark J. G., Hodson W. A., Schmidt R. A., Madtes D. K. Expression of transforming growth factor-alpha in mid-gestation human fetal lung. Am J Respir Cell Mol Biol. 1993 Mar;8(3):266–272. doi: 10.1165/ajrcmb/8.3.266. [DOI] [PubMed] [Google Scholar]
  42. Tomic M., Jiang C. K., Epstein H. S., Freedberg I. M., Samuels H. H., Blumenberg M. Nuclear receptors for retinoic acid and thyroid hormone regulate transcription of keratin genes. Cell Regul. 1990 Nov;1(12):965–973. doi: 10.1091/mbc.1.12.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. VALENTINE E. H. Squamous metaplasia of the bronchus; a study of metaplastic changes occurring in the epithelium of the major bronchi in cancerous and noncancerous cases. Cancer. 1957 Mar-Apr;10(2):272–279. doi: 10.1002/1097-0142(195703/04)10:2<272::aid-cncr2820100209>3.0.co;2-l. [DOI] [PubMed] [Google Scholar]
  44. Vaughan T. J., James P. S., Pascall J. C., Brown K. D. Expression of the genes for TGF alpha, EGF and the EGF receptor during early pig development. Development. 1992 Nov;116(3):663–669. doi: 10.1242/dev.116.3.663. [DOI] [PubMed] [Google Scholar]
  45. Wald N., Idle M., Boreham J., Bailey A. Low serum-vitamin-A and subsequent risk of cancer. Preliminary results of a prospective study. Lancet. 1980 Oct 18;2(8199):813–815. doi: 10.1016/s0140-6736(80)90169-5. [DOI] [PubMed] [Google Scholar]
  46. Wang A. M., Doyle M. V., Mark D. F. Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9717–9721. doi: 10.1073/pnas.86.24.9717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wong Y. C., Buck R. C. An electron microscopic study of metaplasia of the rat tracheal epithelium in vitamin A deficiency. Lab Invest. 1971 Jan;24(1):55–66. [PubMed] [Google Scholar]
  48. Wu R., Smith D. Continuous multiplication of rabbit tracheal epithelial cells in a defined, hormone-supplemented medium. In Vitro. 1982 Sep;18(9):800–812. doi: 10.1007/BF02796504. [DOI] [PubMed] [Google Scholar]
  49. Yasui W., Ji Z. Q., Kuniyasu H., Ayhan A., Yokozaki H., Ito H., Tahara E. Expression of transforming growth factor alpha in human tissues: immunohistochemical study and northern blot analysis. Virchows Arch A Pathol Anat Histopathol. 1992;421(6):513–519. doi: 10.1007/BF01606881. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES