Abstract
Transforming growth factors (TGFs) were initially isolated from the conditioned medium of transformed cell lines through their ability to stimulate anchorage-dependent cells to form colonies in soft agar (1,2). The ability to proliferate in an anchorage-independent manner is still one of the best in vitro correlates with tumorigenicity. Subsequent studies demonstrated that the growth-promoting activity in the conditioned medium consisted of two unique peptides, TGF-α and TGF-β (3–5). Depending on the indicator cell line used, soft-agar colony growth could occur when TGF-α and TGF-β (i.e., NRK cells) or TGF-β alone (i.e., AKR-2B cells) were added to the serum-containing medium (6,7). This review will focus on TGF-β and cellular systems capable of responding in vitro to its growth modulatory activity independent of additional factors.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
DeLarco J. E. and Todaro G. J. (1978) Growth factors from murine sarcoma virus-transformed cells. Proc. Natl. Acad. Sci. USA 75, 4001–4005.
Roberts A. B., Lamb L. C., Newton D. L., Sporn M. B., DeLarco J. E., and Todaro G. J. (1980) Transforming growth factors: Isolation of polypeptides from virally and chemically transformed cells by acid/ethanol extraction. Proc. Natl. Acad. Sci. USA 77, 3494–3498.
Anzano M. A., Roberts A. B., Smith J. M., Sporn M. B., and De Larco J. E. (1983) Sarcoma growth factor from conditioned medium of virally transformed cells is composed of both type alpha and type beta transforming growth factors. Proc. Natl. Acad. Sci. USA 80, 6264–6268.
Moses H. L., Branum E. L., Proper J. A., and Robinson R. A. (1981) Transforming growth factor production by chemically transformed cells. Cancer Res. 41, 2842–2848.
Roberts A. B., Anzano M. A., Lamb L. C., Smith J. M., and Sporn M. B. (1981) New class of transforming growth factors potentiated by epidermal growth factors: Isolation from non-neoplastic tissues. Proc. Natl. Acad. Sci. USA 78, 5339–5343.
Moses H. L., Tucker R. F., Leof E. B., Coffey R. J., Halper J., and Shipley G. D. (1985) Type β transforming growth factor is a growth stimulator and a growth inhibitor. Cancer Cells 3, 65–71.
Roberts A. B., Anzano M. A., Lamb L. C., Smith J. M., Frolik C. A., Marquardt H., Todaro G. J., and Sporn M. B. (1982) Isolation from murine sarcoma cells of novel transforming growth factors potentiated by EGF. Nature 295, 417–419.
Massagué J., Hata A., and Liu F. (1997) TGF-beta signalling through the Smad pathway. Trends Cell Biol. 7, 187–192.
Moses H. L. and Serra R. (1996) Regulation of differentiation by TGF-β. Curr. Opin. Gen. Dev. 6, 581–586.
Hoodless P. A. and Wrana J. L. (1998) Mechanism and function of signaling by the TGF-beta superfamily. Curr. Top. Microbiol. Immunol. 228, 235–272.
Massagué J., Attisano L., and Wrana J. (1994) The TGF beta family and its composite receptors. Trends Cell Biol. 4, 172–178.
Letterio J. J. and Roberts A. B. (1998) Regulation of immune responses by TGF-beta. Annu. Rev. Immunol. 16, 137–161.
Letterio J. J. and Roberts A. B. (1996) Transforming growth factor-b1 deficient mice: identification of isoform specific activities in vivo. J. Leukoc. Biol. 59, 769–774.
Bottinger E. P., Letterio J. J., and Roberts A. B. (1997) Biology of TGF-beta in knockout and transgenic mouse models. Kidney Int. 51, 1355–1360.
Chegini N. (1997) The role of growth factors in peritoneal healing: transforming growth factor beta (TGF-beta). Eur. J. Surg.-Suppl. 577, 17–23.
O’Kane S. and Ferguson M. W. (1997) Transforming growth factor betas and wound healing. Int. J. Biochem. Cell Biol. 29, 63–78.
Franklin T. J. (1997) Therapeutic approaches to organ fibrosis. Int. J. Biochem. Cell Biol. 29, 79–89.
Bitzer M., Sterzel R. B., and Bottinger E. P. (1998) Transforming growth factor-beta in renal disease. Kidney Blood Press. Res. 21, 1–12.
Shipley G. D., Tucker R. F., and Moses H. L. (1985) Type beta transforming growth factor/growth inhibitor stimulates entry of monolayer cultures of AKR-2B cells into S phase after a prolonged prereplicative interval. Proc. Natl. Acad. Sci. USA 82, 4147–4151.
Leof E. B., Proper J. A., Goustin A. S., Shipley G. D., DiCorleto P. E., and Moses H. L. (1986) Induction of c-sis mRNA and activity similar to platelet-derived growth factor by transforming growth factor beta: a proposed model for indirect mitogenesis involving autocrine activity. Proc Natl. Acad. Sci. USA 83, 2453–2457.
Howe P. H., Draetta G., and Leof E. B. (1991) Transforming growth factor beta 1 inhibition of p34cdc2 phosphorylation and histone Hl kinase activity is associated with Gl/S-phase growth arrest. Mol. Cell. Biol 11, 1185–1194.
Cristofalo V. J. and Charpentier R. (1980) A standard procedure for cultivating human diploid fibroblastlike cells to study cellular aging. J. Tissue Cult. Methods 6, 117–121.
Laiho M., Decaprio J. A., Ludlow J. W., Livingston D. M., and Massagué J. (1990) Growth inhibition by TGF-β linked to suppression of retinoblastoma protein phosphorylation. Cell 62, 175–185.
Pietenpol J. A., Stein R. W., Moran E., Yaciuk P., Schlegel R., Lyons R. M., Pittelkow M. R., Munger K., Howley P. M., and Moses H. L. (1990) TGF-beta 1 inhibition of c-myc transcription and growth in keratinocytes is abrogated by viral transforming proteins with pRB binding domains. Cell 6, 777–785.
Shipley G. D. and Ham R. G. (1981) Improved medium and culture conditions for clonal growth with minimal serum protein and for enhanced serum-free survival of Swiss 3T3 cells. In Vitro 17, 656–670.
Leof E. B., Wharton W., Van Wyk J. J., and Pledger W. J. (1982) Epidermal growth factor (EGF) and somatomedin C regulate G1 progression in competent BALB/c-3T3 cells. Exp. Cell Res. 141, 107–115.
Tice D. A., Biscardi J. S., Nickles A. L., and Parsons S. J. (1999) Mechanism of biological synergy between cellular Src and epidermal growth factor receptor. Proc. Natl Acad. Sci USA 96, 1415–1420.
Hanks S. K. and Polte T. R. (1997) Signaling through focal adhesion kinase. Bioessays 19, 137–145.
Weisberg E., Sattler M., Ewaniuk d. S., and Salgia R. (1997) Role of focal adhesion proteins in signal transduction and oncogenesis. Crit. Rev. Oncog. 8, 343–358.
Leof E. B., Proper J. A., Getz M. J., and Moses H. L. (1986) Transforming growth factor type beta regulation of actin mRNA. J. Cell. Physiol. 127, 83–88.
Ignotz R. A. and Massagué J. (1987) Cell adhesion protein receptors as targets for transforming growth factor-beta action. Cell 51, 189–197.
Massagué J., Cheifetz S., Laiho M., Ralph D. A., Weis F. M., and Zentella A. (1992) Transforming growth factor-beta. Cancer Surveys 12, 81–103.
Shipley G. D., Childs C. B., Volkenant M. E., and Moses H. L. (1984) Differential effects of epidermal growth factor, transforming growth factor, and insulin on DNA and protein synthesis and morphology in serum-free cultures of AKR-2B cells. Cancer Res. 44, 710–716.
Anders R. A. and Leof E. B. (1996) Chimeric granulocyte/macrophage colony-stimulating factor/transforming growth factor-β (TGF-β) receptors define a model system for investigating the role of homomeric and heteromeric receptors in TGF-β signaling. J. Biol. Chem. 271, 21,758–21,766.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Edens, M., Leof, E.B. (2000). In Vitro Assays for Measuring TGFβ Growth Stimulation and Inhibition. In: Howe, P.H. (eds) Transforming Growth Factor-Beta Protocols. Methods in Molecular Biology™, vol 142. Humana Press. https://doi.org/10.1385/1-59259-053-5:1
Download citation
DOI: https://doi.org/10.1385/1-59259-053-5:1
Publisher Name: Humana Press
Print ISBN: 978-0-89603-646-8
Online ISBN: 978-1-59259-053-7
eBook Packages: Springer Protocols