Abstract
Autocrine and paracrine modulation of transforming growth factor expression was assessed in rat intestinal epithelial cell lines designated IEC-6 and IEC-7. Addition of the transforming growth factor alpha (TGF alpha) homologue epidermal growth factor (EGF) to media of subconfluent IEC-6 cells led to autocrine stimulation of TGF alpha expression as well as increased expression of the transforming growth factor beta 1 (TGF beta 1). Increased expression of TGF alpha was maximal between 3 and 6 h after addition of EGF and subsequently declined coincident with increasing level of expression of TGF beta 1, which achieved maximal levels 6 h after addition of EGF and was sustained for more than 12 h. Addition of TGF beta 1 also led to autocrine induction of its own expression coincident with suppression of TGF alpha expression. Addition of TGF beta 1 was associated with increased expression of beta-actin when standardized to a constitutive transcript (GAPDH). Similar responses to addition of EGF and TGF beta 1, were observed in another intestinal epithelial cell line, designated IEC-17. Modulation of expression of TGFs was attenuated when cells were grown on the complex extracellular matrix produced by the Engelbreth-Holm-Swarm tumor (Matrigel), reflecting the baseline induction of TGF beta 1 expression when compared to IEC-6 and IEC-17 cells maintained on plastic. These observations suggest that expression of TGFs is controlled by autocrine mechanisms in intestinal epithelial cell lines and proliferation stimulated by TGF alpha may be initially self-reinforcing but ultimately downregulated by induction of TGF beta 1.
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- Anzano M. A., Rieman D., Prichett W., Bowen-Pope D. F., Greig R. Growth factor production by human colon carcinoma cell lines. Cancer Res. 1989 Jun 1;49(11):2898–2904. [PubMed] [Google Scholar]
- Barnard J. A., Beauchamp R. D., Coffey R. J., Moses H. L. Regulation of intestinal epithelial cell growth by transforming growth factor type beta. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1578–1582. doi: 10.1073/pnas.86.5.1578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boyd D., Florent G., Chakrabarty S., Brattain D., Brattain M. G. Alterations of the biological characteristics of a colon carcinoma cell line by colon-derived substrata material. Cancer Res. 1988 May 15;48(10):2825–2831. [PubMed] [Google Scholar]
- Brachmann R., Lindquist P. B., Nagashima M., Kohr W., Lipari T., Napier M., Derynck R. Transmembrane TGF-alpha precursors activate EGF/TGF-alpha receptors. Cell. 1989 Feb 24;56(4):691–700. doi: 10.1016/0092-8674(89)90591-6. [DOI] [PubMed] [Google Scholar]
- Carroll K. M., Wong T. T., Drabik D. L., Chang E. B. Differentiation of rat small intestinal epithelial cells by extracellular matrix. Am J Physiol. 1988 Mar;254(3 Pt 1):G355–G360. doi: 10.1152/ajpgi.1988.254.3.G355. [DOI] [PubMed] [Google Scholar]
- 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]
- Coffey R. J., Jr, Goustin A. S., Soderquist A. M., Shipley G. D., Wolfshohl J., Carpenter G., Moses H. L. Transforming growth factor alpha and beta expression in human colon cancer lines: implications for an autocrine model. Cancer Res. 1987 Sep 1;47(17):4590–4594. [PubMed] [Google Scholar]
- Coffey R. J., Jr, Shipley G. D., Moses H. L. Production of transforming growth factors by human colon cancer lines. Cancer Res. 1986 Mar;46(3):1164–1169. [PubMed] [Google Scholar]
- Coffey R. J., Jr, Sipes N. J., Bascom C. C., Graves-Deal R., Pennington C. Y., Weissman B. E., Moses H. L. Growth modulation of mouse keratinocytes by transforming growth factors. Cancer Res. 1988 Mar 15;48(6):1596–1602. [PubMed] [Google Scholar]
- Derynck R., Jarrett J. A., Chen E. Y., Eaton D. H., Bell J. R., Assoian R. K., Roberts A. B., Sporn M. B., Goeddel D. V. Human transforming growth factor-beta complementary DNA sequence and expression in normal and transformed cells. Nature. 1985 Aug 22;316(6030):701–705. doi: 10.1038/316701a0. [DOI] [PubMed] [Google Scholar]
- Gonnella P. A., Siminoski K., Murphy R. A., Neutra M. R. Transepithelial transport of epidermal growth factor by absorptive cells of suckling rat ileum. J Clin Invest. 1987 Jul;80(1):22–32. doi: 10.1172/JCI113051. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hahn U., Stallmach A., Hahn E. G., Riecken E. O. Basement membrane components are potent promoters of rat intestinal epithelial cell differentiation in vitro. Gastroenterology. 1990 Feb;98(2):322–335. doi: 10.1016/0016-5085(90)90821-h. [DOI] [PubMed] [Google Scholar]
- Hoosein N. M., Brattain D. E., McKnight M. K., Levine A. E., Brattain M. G. Characterization of the inhibitory effects of transforming growth factor-beta on a human colon carcinoma cell line. Cancer Res. 1987 Jun 1;47(11):2950–2954. [PubMed] [Google Scholar]
- Kleinman H. K., McGarvey M. L., Liotta L. A., Robey P. G., Tryggvason K., Martin G. R. Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma. Biochemistry. 1982 Nov 23;21(24):6188–6193. doi: 10.1021/bi00267a025. [DOI] [PubMed] [Google Scholar]
- Koyama S. Y., Podolsky D. K. Differential expression of transforming growth factors alpha and beta in rat intestinal epithelial cells. J Clin Invest. 1989 May;83(5):1768–1773. doi: 10.1172/JCI114080. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurokowa M., Lynch K., Podolsky D. K. Effects of growth factors on an intestinal epithelial cell line: transforming growth factor beta inhibits proliferation and stimulates differentiation. Biochem Biophys Res Commun. 1987 Feb 13;142(3):775–782. doi: 10.1016/0006-291x(87)91481-1. [DOI] [PubMed] [Google Scholar]
- Massagué J., Cheifetz S., Ignotz R. A., Boyd F. T. Multiple type-beta transforming growth factors and their receptors. J Cell Physiol Suppl. 1987;Suppl 5:43–47. doi: 10.1002/jcp.1041330409. [DOI] [PubMed] [Google Scholar]
- 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. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moses H. L., Coffey R. J., Jr, Leof E. B., Lyons R. M., Keski-Oja J. Transforming growth factor beta regulation of cell proliferation. J Cell Physiol Suppl. 1987;Suppl 5:1–7. doi: 10.1002/jcp.1041330403. [DOI] [PubMed] [Google Scholar]
- Mulder K. M., Levine A. E., Hernandez X., McKnight M. K., Brattain D. E., Brattain M. G. Modulation of c-myc by transforming growth factor-beta in human colon carcinoma cells. Biochem Biophys Res Commun. 1988 Jan 29;150(2):711–716. doi: 10.1016/0006-291x(88)90449-4. [DOI] [PubMed] [Google Scholar]
- Mulder K. M., Ramey M. K., Hoosein N. M., Levine A. E., Hinshaw X. H., Brattain D. E., Brattain M. G. Characterization of transforming growth factor-beta-resistant subclones isolated from a transforming growth factor-beta-sensitive human colon carcinoma cell line. Cancer Res. 1988 Dec 15;48(24 Pt 1):7120–7125. [PubMed] [Google Scholar]
- Ménard D., Arsenault P., Pothier P. Biologic effects of epidermal growth factor in human fetal jejunum. Gastroenterology. 1988 Mar;94(3):656–663. doi: 10.1016/0016-5085(88)90236-3. [DOI] [PubMed] [Google Scholar]
- Podolsky D. K., Pleskow D. K., Jafari H. Latent transformed growth-inhibiting factor in human malignant effusions. Cancer Res. 1988 Jan 15;48(2):418–424. [PubMed] [Google Scholar]
- Quaroni A., Wands J., Trelstad R. L., Isselbacher K. J. Epithelioid cell cultures from rat small intestine. Characterization by morphologic and immunologic criteria. J Cell Biol. 1979 Feb;80(2):248–265. doi: 10.1083/jcb.80.2.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scarpa S., D'Orazi G., Modesti A., Valia S., Zucco F., Stammati A. Expression and synthesis of fibronectin and laminin by an intestinal epithelial cell line. Tissue Cell. 1988;20(3):305–312. doi: 10.1016/0040-8166(88)90066-3. [DOI] [PubMed] [Google Scholar]
- Scheving L. A., Shiurba R. A., Nguyen T. D., Gray G. M. Epidermal growth factor receptor of the intestinal enterocyte. Localization to laterobasal but not brush border membrane. J Biol Chem. 1989 Jan 25;264(3):1735–1741. [PubMed] [Google Scholar]
- Seo M. K., Lynch K. E., Podolsky D. K. Multiplicity of transforming growth factors in human malignant effusions. Cancer Res. 1988 Apr 1;48(7):1792–1797. [PubMed] [Google Scholar]
- Stallmach A., Hahn U., Merker H. J., Hahn E. G., Riecken E. O. Differentiation of rat intestinal epithelial cells is induced by organotypic mesenchymal cells in vitro. Gut. 1989 Jul;30(7):959–970. doi: 10.1136/gut.30.7.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weaver L. T., Gonnella P. A., Israel E. J., Walker W. A. Uptake and transport of epidermal growth factor by the small intestinal epithelium of the fetal rat. Gastroenterology. 1990 Apr;98(4):828–837. doi: 10.1016/0016-5085(90)90004-k. [DOI] [PubMed] [Google Scholar]
- Winkler M. E., O'Connor L., Winget M., Fendly B. Epidermal growth factor and transforming growth factor alpha bind differently to the epidermal growth factor receptor. Biochemistry. 1989 Jul 25;28(15):6373–6378. doi: 10.1021/bi00441a033. [DOI] [PubMed] [Google Scholar]
- Wong S. T., Winchell L. F., McCune B. K., Earp H. S., Teixidó J., Massagué J., Herman B., Lee D. C. The TGF-alpha precursor expressed on the cell surface binds to the EGF receptor on adjacent cells, leading to signal transduction. Cell. 1989 Feb 10;56(3):495–506. doi: 10.1016/0092-8674(89)90252-3. [DOI] [PubMed] [Google Scholar]