Growth stimulation of human breast cancer cells with anti-transforming growth factor beta antibodies

evidence for negative autocrine regulation by transforming growth factor beta.

C. L. Arteaga, R. J. Coffey, T. C. Dugger, C. M. McCutchen, H. L. Moses, R. M. Lyons

Research output: Contribution to journalArticle

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Abstract

Exogenous TGF beta inhibits the proliferation of human breast cancer cells in vitro. These cells synthesize and secrete TGF beta into their medium predominantly in a latent form. With neutralizing antibodies against native, biologically active TGF beta (278ab and 282ab), we have examined whether HS578T and MDA-231 breast cancer cells utilize their endogenous TGF beta for growth regulation. Low levels of TGF beta activity were detectable in conditioned medium from confluent monolayers of both cell lines in the absence of acid or protease treatment as measured by radioreceptor assay. When added to subconfluent monolayers of the respective cell line, this untreated conditioned medium inhibited DNA synthesis and cell proliferation. This inhibition was blocked by anti-TGF beta antibodies, whereas nonimmune rabbit IgG had no effect. Similar to exogenous TGF beta 1, this conditioned medium induced a dose-dependent increase in steady-state TGF beta 1 mRNA levels when added to subconfluent HS578T cells; this increase was blocked by the 278ab. Consistent with the above, preincubation of either cell line with anti-TGF beta antibodies increased subsequent specific binding of 125I-TGF beta to cell surface receptors without changing binding affinity. Addition of 278ab to quiescent HS578T or MDA-231 cells induced a dose-dependent increase in [3H]thymidine incorporation. Both antibodies stimulated cell proliferation in serum-free medium and anchorage-independent growth of both cell lines. Finally, incubation of HS578T cells with 278ab under serum-free conditions decreased the basal level of TGF beta 1 message expression. These data indicate that cultured human breast cancer cells utilize endogenously produced TGF beta as an autocrine negative growth regulator.

Original languageEnglish (US)
Pages (from-to)367-374
Number of pages8
JournalCell growth & differentiation : the molecular biology journal of the American Association for Cancer Research
Volume1
Issue number8
StatePublished - Aug 1 1990

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Transforming Growth Factor beta
Breast Neoplasms
Antibodies
Growth
Transforming Growth Factor beta1
Conditioned Culture Medium
Cell Line
Cell Proliferation
Radioligand Assay
Serum-Free Culture Media
Cell Surface Receptors
Neutralizing Antibodies
Thymidine
Peptide Hydrolases
Immunoglobulin G
Rabbits
Messenger RNA
Acids
DNA
Serum

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

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title = "Growth stimulation of human breast cancer cells with anti-transforming growth factor beta antibodies: evidence for negative autocrine regulation by transforming growth factor beta.",
abstract = "Exogenous TGF beta inhibits the proliferation of human breast cancer cells in vitro. These cells synthesize and secrete TGF beta into their medium predominantly in a latent form. With neutralizing antibodies against native, biologically active TGF beta (278ab and 282ab), we have examined whether HS578T and MDA-231 breast cancer cells utilize their endogenous TGF beta for growth regulation. Low levels of TGF beta activity were detectable in conditioned medium from confluent monolayers of both cell lines in the absence of acid or protease treatment as measured by radioreceptor assay. When added to subconfluent monolayers of the respective cell line, this untreated conditioned medium inhibited DNA synthesis and cell proliferation. This inhibition was blocked by anti-TGF beta antibodies, whereas nonimmune rabbit IgG had no effect. Similar to exogenous TGF beta 1, this conditioned medium induced a dose-dependent increase in steady-state TGF beta 1 mRNA levels when added to subconfluent HS578T cells; this increase was blocked by the 278ab. Consistent with the above, preincubation of either cell line with anti-TGF beta antibodies increased subsequent specific binding of 125I-TGF beta to cell surface receptors without changing binding affinity. Addition of 278ab to quiescent HS578T or MDA-231 cells induced a dose-dependent increase in [3H]thymidine incorporation. Both antibodies stimulated cell proliferation in serum-free medium and anchorage-independent growth of both cell lines. Finally, incubation of HS578T cells with 278ab under serum-free conditions decreased the basal level of TGF beta 1 message expression. These data indicate that cultured human breast cancer cells utilize endogenously produced TGF beta as an autocrine negative growth regulator.",
author = "Arteaga, {C. L.} and Coffey, {R. J.} and Dugger, {T. C.} and McCutchen, {C. M.} and Moses, {H. L.} and Lyons, {R. M.}",
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T2 - evidence for negative autocrine regulation by transforming growth factor beta.

AU - Arteaga, C. L.

AU - Coffey, R. J.

AU - Dugger, T. C.

AU - McCutchen, C. M.

AU - Moses, H. L.

AU - Lyons, R. M.

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N2 - Exogenous TGF beta inhibits the proliferation of human breast cancer cells in vitro. These cells synthesize and secrete TGF beta into their medium predominantly in a latent form. With neutralizing antibodies against native, biologically active TGF beta (278ab and 282ab), we have examined whether HS578T and MDA-231 breast cancer cells utilize their endogenous TGF beta for growth regulation. Low levels of TGF beta activity were detectable in conditioned medium from confluent monolayers of both cell lines in the absence of acid or protease treatment as measured by radioreceptor assay. When added to subconfluent monolayers of the respective cell line, this untreated conditioned medium inhibited DNA synthesis and cell proliferation. This inhibition was blocked by anti-TGF beta antibodies, whereas nonimmune rabbit IgG had no effect. Similar to exogenous TGF beta 1, this conditioned medium induced a dose-dependent increase in steady-state TGF beta 1 mRNA levels when added to subconfluent HS578T cells; this increase was blocked by the 278ab. Consistent with the above, preincubation of either cell line with anti-TGF beta antibodies increased subsequent specific binding of 125I-TGF beta to cell surface receptors without changing binding affinity. Addition of 278ab to quiescent HS578T or MDA-231 cells induced a dose-dependent increase in [3H]thymidine incorporation. Both antibodies stimulated cell proliferation in serum-free medium and anchorage-independent growth of both cell lines. Finally, incubation of HS578T cells with 278ab under serum-free conditions decreased the basal level of TGF beta 1 message expression. These data indicate that cultured human breast cancer cells utilize endogenously produced TGF beta as an autocrine negative growth regulator.

AB - Exogenous TGF beta inhibits the proliferation of human breast cancer cells in vitro. These cells synthesize and secrete TGF beta into their medium predominantly in a latent form. With neutralizing antibodies against native, biologically active TGF beta (278ab and 282ab), we have examined whether HS578T and MDA-231 breast cancer cells utilize their endogenous TGF beta for growth regulation. Low levels of TGF beta activity were detectable in conditioned medium from confluent monolayers of both cell lines in the absence of acid or protease treatment as measured by radioreceptor assay. When added to subconfluent monolayers of the respective cell line, this untreated conditioned medium inhibited DNA synthesis and cell proliferation. This inhibition was blocked by anti-TGF beta antibodies, whereas nonimmune rabbit IgG had no effect. Similar to exogenous TGF beta 1, this conditioned medium induced a dose-dependent increase in steady-state TGF beta 1 mRNA levels when added to subconfluent HS578T cells; this increase was blocked by the 278ab. Consistent with the above, preincubation of either cell line with anti-TGF beta antibodies increased subsequent specific binding of 125I-TGF beta to cell surface receptors without changing binding affinity. Addition of 278ab to quiescent HS578T or MDA-231 cells induced a dose-dependent increase in [3H]thymidine incorporation. Both antibodies stimulated cell proliferation in serum-free medium and anchorage-independent growth of both cell lines. Finally, incubation of HS578T cells with 278ab under serum-free conditions decreased the basal level of TGF beta 1 message expression. These data indicate that cultured human breast cancer cells utilize endogenously produced TGF beta as an autocrine negative growth regulator.

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