Abstract
Acquisition of cell motility is often correlated with the malignant progression of a transformed cell. To investigate some of the mechanisms involved in the development of a migratory state, we transfected the NBTII rat carcinoma cell line, which forms stationary epithelial clusters in culture, with the gene encoding human transforming growth factor alpha (TGF alpha). Expression of TGF alpha in NBTII cells resulted in cells of motile and vimentin-positive phenotype with internalized desmosomal components, analogous to the treatment of cells with exogenous TGF alpha. The clones expressed a 5.2-kb TGF alpha message and synthesized an 18-kDa form of TGF alpha. Supernatants of TGF alpha-producing clones induced the internalization of desmosomal components, the production of vimentin, and increased motility in untransfected epithelial NBTII cells, indicating that the factor produced by the clones was in a biologically active form. TGF alpha-producing clones secreted significant levels of a 95-kDa gelatinolytic metal-loproteinase, virtually absent in untransfected cell supernatants. In contrast, levels of inhibitors of metalloproteinases and of a plasminogen activator were similar in untransfected and TGF alpha-transfected NBTII cells. These results suggest that expression of TGF alpha in an epithelial tumor cell results in the development of a motile, fibroblast-like phenotype with matrix-degrading potential, which could result in a more aggressive tumor in vivo.
Full text
PDF











Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barrandon Y., Green H. Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. Cell. 1987 Sep 25;50(7):1131–1137. doi: 10.1016/0092-8674(87)90179-6. [DOI] [PubMed] [Google Scholar]
- Birkedal-Hansen H., Taylor R. E. Detergent-activation of latent collagenase and resolution of its component molecules. Biochem Biophys Res Commun. 1982 Aug 31;107(4):1173–1178. doi: 10.1016/s0006-291x(82)80120-4. [DOI] [PubMed] [Google Scholar]
- Boyer B., Tucker G. C., Vallés A. M., Franke W. W., Thiery J. P. Rearrangements of desmosomal and cytoskeletal proteins during the transition from epithelial to fibroblastoid organization in cultured rat bladder carcinoma cells. J Cell Biol. 1989 Oct;109(4 Pt 1):1495–1509. doi: 10.1083/jcb.109.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bringman T. S., Lindquist P. B., Derynck R. Different transforming growth factor-alpha species are derived from a glycosylated and palmitoylated transmembrane precursor. Cell. 1987 Feb 13;48(3):429–440. doi: 10.1016/0092-8674(87)90194-2. [DOI] [PubMed] [Google Scholar]
- Cawston T. E., Galloway W. A., Mercer E., Murphy G., Reynolds J. J. Purification of rabbit bone inhibitor of collagenase. Biochem J. 1981 Apr 1;195(1):159–165. doi: 10.1042/bj1950159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chua C. C., Geiman D. E., Keller G. H., Ladda R. L. Induction of collagenase secretion in human fibroblast cultures by growth promoting factors. J Biol Chem. 1985 May 10;260(9):5213–5216. [PubMed] [Google Scholar]
- Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. A new dominant hybrid selective marker for higher eukaryotic cells. J Mol Biol. 1981 Jul 25;150(1):1–14. doi: 10.1016/0022-2836(81)90321-1. [DOI] [PubMed] [Google Scholar]
- De Clerck Y. A., Yean T. D., Ratzkin B. J., Lu H. S., Langley K. E. Purification and characterization of two related but distinct metalloproteinase inhibitors secreted by bovine aortic endothelial cells. J Biol Chem. 1989 Oct 15;264(29):17445–17453. [PubMed] [Google Scholar]
- Derynck R. Transforming growth factor alpha. Cell. 1988 Aug 26;54(5):593–595. doi: 10.1016/s0092-8674(88)80001-1. [DOI] [PubMed] [Google Scholar]
- Dubeau L., Jones P. A., Rideout W. M., 3rd, Laug W. E. Differential regulation of plasminogen activators by epidermal growth factor in normal and neoplastic human urothelium. Cancer Res. 1988 Oct 1;48(19):5552–5556. [PubMed] [Google Scholar]
- Edwards D. R., Murphy G., Reynolds J. J., Whitham S. E., Docherty A. J., Angel P., Heath J. K. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor. EMBO J. 1987 Jul;6(7):1899–1904. doi: 10.1002/j.1460-2075.1987.tb02449.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
- Finzi E., Fleming T., Segatto O., Pennington C. Y., Bringman T. S., Derynck R., Aaronson S. A. The human transforming growth factor type alpha coding sequence is not a direct-acting oncogene when overexpressed in NIH 3T3 cells. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3733–3737. doi: 10.1073/pnas.84.11.3733. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finzi E., Kilkenny A., Strickland J. E., Balaschak M., Bringman T., Derynck R., Aaronson S., Yuspa S. H. TGF alpha stimulates growth of skin papillomas by autocrine and paracrine mechanisms but does not cause neoplastic progression. Mol Carcinog. 1988;1(1):7–12. doi: 10.1002/mc.2940010105. [DOI] [PubMed] [Google Scholar]
- Fort P., Marty L., Piechaczyk M., el Sabrouty S., Dani C., Jeanteur P., Blanchard J. M. Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res. 1985 Mar 11;13(5):1431–1442. doi: 10.1093/nar/13.5.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gavrilovic J., Hembry R. M., Reynolds J. J., Murphy G. Tissue inhibitor of metalloproteinases (TIMP) regulates extracellular type I collagen degradation by chondrocytes and endothelial cells. J Cell Sci. 1987 Mar;87(Pt 2):357–362. doi: 10.1242/jcs.87.2.357. [DOI] [PubMed] [Google Scholar]
- Gherardi E., Gray J., Stoker M., Perryman M., Furlong R. Purification of scatter factor, a fibroblast-derived basic protein that modulates epithelial interactions and movement. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5844–5848. doi: 10.1073/pnas.86.15.5844. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
- Grey A. M., Schor A. M., Rushton G., Ellis I., Schor S. L. Purification of the migration stimulating factor produced by fetal and breast cancer patient fibroblasts. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2438–2442. doi: 10.1073/pnas.86.7.2438. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herron G. S., Banda M. J., Clark E. J., Gavrilovic J., Werb Z. Secretion of metalloproteinases by stimulated capillary endothelial cells. II. Expression of collagenase and stromelysin activities is regulated by endogenous inhibitors. J Biol Chem. 1986 Feb 25;261(6):2814–2818. [PubMed] [Google Scholar]
- Heussen C., Dowdle E. B. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem. 1980 Feb;102(1):196–202. doi: 10.1016/0003-2697(80)90338-3. [DOI] [PubMed] [Google Scholar]
- Ignotz R. A., Kelly B., Davis R. J., Massagué J. Biologically active precursor for transforming growth factor type alpha, released by retrovirally transformed cells. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6307–6311. doi: 10.1073/pnas.83.17.6307. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- Lee D. C., Rose T. M., Webb N. R., Todaro G. J. Cloning and sequence analysis of a cDNA for rat transforming growth factor-alpha. Nature. 1985 Feb 7;313(6002):489–491. doi: 10.1038/313489a0. [DOI] [PubMed] [Google Scholar]
- Lee P. L., Johnson D. E., Cousens L. S., Fried V. A., Williams L. T. Purification and complementary DNA cloning of a receptor for basic fibroblast growth factor. Science. 1989 Jul 7;245(4913):57–60. doi: 10.1126/science.2544996. [DOI] [PubMed] [Google Scholar]
- Liotta L. A., Mandler R., Murano G., Katz D. A., Gordon R. K., Chiang P. K., Schiffmann E. Tumor cell autocrine motility factor. Proc Natl Acad Sci U S A. 1986 May;83(10):3302–3306. doi: 10.1073/pnas.83.10.3302. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liotta L. A. Tumor invasion and metastases: role of the basement membrane. Warner-Lambert Parke-Davis Award lecture. Am J Pathol. 1984 Dec;117(3):339–348. [PMC free article] [PubMed] [Google Scholar]
- Luetteke N. C., Michalopoulos G. K., Teixidó J., Gilmore R., Massagué J., Lee D. C. Characterization of high molecular weight transforming growth factor alpha produced by rat hepatocellular carcinoma cells. Biochemistry. 1988 Aug 23;27(17):6487–6494. doi: 10.1021/bi00417a043. [DOI] [PubMed] [Google Scholar]
- Matrisian L. M., Glaichenhaus N., Gesnel M. C., Breathnach R. Epidermal growth factor and oncogenes induce transcription of the same cellular mRNA in rat fibroblasts. EMBO J. 1985 Jun;4(6):1435–1440. doi: 10.1002/j.1460-2075.1985.tb03799.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murphy G., Cawston T. E., Reynolds J. J. An inhibitor of collagenase from human amniotic fluid. Purification, characterization and action on metalloproteinases. Biochem J. 1981 Apr 1;195(1):167–170. doi: 10.1042/bj1950167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murphy G., McAlpine C. G., Poll C. T., Reynolds J. J. Purification and characterization of a bone metalloproteinase that degrades gelatin and types IV and V collagen. Biochim Biophys Acta. 1985 Sep 20;831(1):49–58. doi: 10.1016/0167-4838(85)90148-7. [DOI] [PubMed] [Google Scholar]
- Presta M., Moscatelli D., Joseph-Silverstein J., Rifkin D. B. Purification from a human hepatoma cell line of a basic fibroblast growth factor-like molecule that stimulates capillary endothelial cell plasminogen activator production, DNA synthesis, and migration. Mol Cell Biol. 1986 Nov;6(11):4060–4066. doi: 10.1128/mcb.6.11.4060. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosen E. M., Meromsky L., Setter E., Vinter D. W., Goldberg I. D. Smooth muscle-derived factor stimulates mobility of human tumor cells. Invasion Metastasis. 1990;10(1):49–64. [PubMed] [Google Scholar]
- Rosenthal A., Lindquist P. B., Bringman T. S., Goeddel D. V., Derynck R. Expression in rat fibroblasts of a human transforming growth factor-alpha cDNA results in transformation. Cell. 1986 Jul 18;46(2):301–309. doi: 10.1016/0092-8674(86)90747-6. [DOI] [PubMed] [Google Scholar]
- Sato Y., Rifkin D. B. Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis. J Cell Biol. 1988 Sep;107(3):1199–1205. doi: 10.1083/jcb.107.3.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stetler-Stevenson W. G., Krutzsch H. C., Liotta L. A. Tissue inhibitor of metalloproteinase (TIMP-2). A new member of the metalloproteinase inhibitor family. J Biol Chem. 1989 Oct 15;264(29):17374–17378. [PubMed] [Google Scholar]
- Stracke M. L., Kohn E. C., Aznavoorian S. A., Wilson L. L., Salomon D., Krutzsch H. C., Liotta L. A., Schiffmann E. Insulin-like growth factors stimulate chemotaxis in human melanoma cells. Biochem Biophys Res Commun. 1988 Jun 30;153(3):1076–1083. doi: 10.1016/s0006-291x(88)81338-x. [DOI] [PubMed] [Google Scholar]
- Toyoshima K., Ito N., Hiasa Y., Kamamoto Y., Makiura S. Tissue culture of urinary bladder tumor induced in a rat by N-butyl-N-(-4-hydroxybutyl)nitrosamine: establishment of cell line, Nara Bladder Tumor II. J Natl Cancer Inst. 1971 Nov;47(5):979–985. [PubMed] [Google Scholar]
- Tsuboi R., Sato Y., Rifkin D. B. Correlation of cell migration, cell invasion, receptor number, proteinase production, and basic fibroblast growth factor levels in endothelial cells. J Cell Biol. 1990 Feb;110(2):511–517. doi: 10.1083/jcb.110.2.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tucker G. C., Boyer B., Gavrilovic J., Emonard H., Thiery J. P. Collagen-mediated dispersion of NBT-II rat bladder carcinoma cells. Cancer Res. 1990 Jan 1;50(1):129–137. [PubMed] [Google Scholar]
- Vallés A. M., Boyer B., Badet J., Tucker G. C., Barritault D., Thiery J. P. Acidic fibroblast growth factor is a modulator of epithelial plasticity in a rat bladder carcinoma cell line. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1124–1128. doi: 10.1073/pnas.87.3.1124. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilhelm S. M., Collier I. E., Marmer B. L., Eisen A. Z., Grant G. A., Goldberg G. I. SV40-transformed human lung fibroblasts secrete a 92-kDa type IV collagenase which is identical to that secreted by normal human macrophages. J Biol Chem. 1989 Oct 15;264(29):17213–17221. [PubMed] [Google Scholar]
- Woolley D. E. Collagenolytic mechanisms in tumor cell invasion. Cancer Metastasis Rev. 1984;3(4):361–372. doi: 10.1007/BF00051460. [DOI] [PubMed] [Google Scholar]
- Yamagata S., Ito Y., Tanaka R., Shimizu S. Gelatinases of metastatic cell lines of murine colonic carcinoma as detected by substrate-gel electrophoresis. Biochem Biophys Res Commun. 1988 Feb 29;151(1):158–162. doi: 10.1016/0006-291x(88)90573-6. [DOI] [PubMed] [Google Scholar]
- Yamagata S., Tanaka R., Ito Y., Shimizu S. Gelatinases of murine metastatic tumor cells. Biochem Biophys Res Commun. 1989 Jan 16;158(1):228–234. doi: 10.1016/s0006-291x(89)80202-5. [DOI] [PubMed] [Google Scholar]