Lack of Correlation between Sensitivity to Growth Inhibition and Receptor Number for Transforming Growth Factor ÃŸin Human Squamous Carcinoma Cell Lines

Four human squamous carcinoma cell lines, SCC-9, SCC-12F, SCC- 15G, and SCC-25, were examined for sensitivity to the growth-inhibitory and differentiation-inducing effects of transforming growth factor ÃŸ (TGF-/3). None of the four cell lines was induced to differentiate, as measured by staining for keratin and by competence of cells to form cross-linked envelopes, by concentrations of TGF-/S as high as 1000 pM. TGF-0 did not inhibit DNA synthesis or proliferation of SCC-12F cells in monolayer culture. Monolayer growth and DNA synthesis of SCC- 15G and SCC-25 cells were markedly inhibited by 10 pM TGF-,8, with maximal inhibition between 10 and 100 P.M. Inhibition of SCC-15G cells was apparent as early as 8 h after addition of TGF-/3, but inhibition of SCC-25 cells was not measurable until 2 days. Growth inhibition of SCC-15G cells was completely reversible, whereas inhibition of SCC-25 cells was irreversible. When growth of cells was measured in a defined medium supplemented with 0.5% fetal bovine serum, TGF-0 inhibited both the monolayer and clonal growth of SCC-15G cells. The monolayer growth, but not the clonal growth, of SCC-25 cells was inhibited. Growth of SCC-9 cells in confluent cultures was slightly inhibited by TGF-/3, while growth in subconfluent cultures was unaffected. All four cell lines, when assayed for binding of 12SI-labeledTGF-0, displayed high-affinity (Kv = 2-44 pM) binding sites. These sites were present in very low numbers (<2000 sites/cell) in SCC-9, SCC-15G, and SCC-25 cells. SCC-12F cells contained 2000-5500 high-affinity sites/cell. Thus, a lack of sensitivity to growth inhibition by TGK-/3 does not necessarily correlate with an inability to bind the growth factor to specific cell surface receptors. The possibility remains that altered sensitivity to TGF-/3 may be due to loss of or changes in the relative proportions of the various TGF-/3 receptor types. TGF-/3 is believed to play a primary role in control of normal cellular growth and differentiation, and the study of cells with defective responses to this growth factor should help to identify some of the critical points within the growth cycle at which malignant cells may escape normal regulatory controls. The differential responses of these four human squamous carcinoma cell lines to TGF-/? should provide a useful model for studying such control mechanisms. properties (8, 18, 19). It has been postu lated that a lack of responsiveness to negative growth control mechanisms such as TGF-0 may lead to the aberrant growth observed in cancer. The purpose of this study was to compare various human squamous cell carcinoma lines with regard to their sensitivity to the growth-inhibitory and differentiation-inducing effects of TGF-/3 and to examine the correlation between this response and the levels of specific TGF-0 receptors in these cells. The results support a model for a complex differentiation pathway with multiple control points in epithelial cells. The carcinoma cell lines used in this study appear to have defects at different points in this multistep pathway and should be useful in eluci dating details of the mechanisms by which cancer cells escape normal growth controls.