TGF-β1 is an autocrine-negative growth regulator of human colon carcinoma FET cells in vivo as revealed by transfection of an antisense expression vector

S. Wu, D. Theodorescu, R. S. Kerbel, J. K.V. Willson, K. M. Mulder, L. E. Humphrey, Michael G Brattain

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Abstract

Transforming growth factor-β1 (TGF-β1) has previously been implicated as a potential negative autocrine or paracrine growth regulator of certain cell types (Arteaga, C. L., R. J. Coffey, Jr., T. C. Dugger, C. M. McCutchen, H. L. Moses, and R. M. Lyons. 1990. Cell Growth and Differ. 1:367-374; Hafez, M. M., D. Infante, S. Winawer, and E. Friedman. 1990. Cell Growth and Differ. 1:617-626; Glick, A. B., K. C. Flanders, D. Danielpour, S. H. Yuspa, and M. B. Sporn. 1989. Cell Regulation. 1:87-97). This is based mainly on experiments assessing the effects of exogenous TGF-β1 or neutralizing antibodies to TGF-β1 on normal or tumor cell proliferation in vitro. However, direct evidence demonstrating such a negative regulation of tumor cell growth in vivo is still lacking. To overcome this problem we have constructed and used an antisense expression vector for TGF-β1 as a means of regulating endogenous TGF-β1 expression in tumor cells. Antisense-transfected FET human colon carcinoma cells showed a fivefold reduction in TGF-β1 mRNA and 15-fold reduction in TGF-β1 secretion. Antisense mRNA was detected in transfected cells by an RNase protection assay. Compared to control cells, cultured antisense-transfected cells showed a reduction in lag phase time rather than a change in doubling time. Cloning efficiencies of transfected cells were four times greater than control cells in anchorage-independent assays. Control cells did not form tumors at 5 x 105 in athymic nude mice. Antisense-transfected cells formed tumors in 40% of animals injected. At higher inocula (1 x 106 cells) antisense-transfected cells formed tumors in 100% of animals injected, but control cells still failed to form tumors. These results show that TGF-β1 acts as a negative growth regulator of human colon carcinoma cells in vivo as well as in vitro. Acquisition of partial or full resistance to such inhibitory effects may therefore contribute to tumor development and progression.

Original languageEnglish (US)
Pages (from-to)187-196
Number of pages10
JournalJournal of Cell Biology
Volume116
Issue number1
DOIs
StatePublished - Jan 1 1992

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Transforming Growth Factors
Transfection
Colon
Carcinoma
Growth
Neoplasms
Nude Mice
Messenger RNA
Cytoprotection
Ribonucleases
Neutralizing Antibodies
Organism Cloning

ASJC Scopus subject areas

  • Cell Biology

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TGF-β1 is an autocrine-negative growth regulator of human colon carcinoma FET cells in vivo as revealed by transfection of an antisense expression vector. / Wu, S.; Theodorescu, D.; Kerbel, R. S.; Willson, J. K.V.; Mulder, K. M.; Humphrey, L. E.; Brattain, Michael G.

In: Journal of Cell Biology, Vol. 116, No. 1, 01.01.1992, p. 187-196.

Research output: Contribution to journalArticle

Wu, S. ; Theodorescu, D. ; Kerbel, R. S. ; Willson, J. K.V. ; Mulder, K. M. ; Humphrey, L. E. ; Brattain, Michael G. / TGF-β1 is an autocrine-negative growth regulator of human colon carcinoma FET cells in vivo as revealed by transfection of an antisense expression vector. In: Journal of Cell Biology. 1992 ; Vol. 116, No. 1. pp. 187-196.
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abstract = "Transforming growth factor-β1 (TGF-β1) has previously been implicated as a potential negative autocrine or paracrine growth regulator of certain cell types (Arteaga, C. L., R. J. Coffey, Jr., T. C. Dugger, C. M. McCutchen, H. L. Moses, and R. M. Lyons. 1990. Cell Growth and Differ. 1:367-374; Hafez, M. M., D. Infante, S. Winawer, and E. Friedman. 1990. Cell Growth and Differ. 1:617-626; Glick, A. B., K. C. Flanders, D. Danielpour, S. H. Yuspa, and M. B. Sporn. 1989. Cell Regulation. 1:87-97). This is based mainly on experiments assessing the effects of exogenous TGF-β1 or neutralizing antibodies to TGF-β1 on normal or tumor cell proliferation in vitro. However, direct evidence demonstrating such a negative regulation of tumor cell growth in vivo is still lacking. To overcome this problem we have constructed and used an antisense expression vector for TGF-β1 as a means of regulating endogenous TGF-β1 expression in tumor cells. Antisense-transfected FET human colon carcinoma cells showed a fivefold reduction in TGF-β1 mRNA and 15-fold reduction in TGF-β1 secretion. Antisense mRNA was detected in transfected cells by an RNase protection assay. Compared to control cells, cultured antisense-transfected cells showed a reduction in lag phase time rather than a change in doubling time. Cloning efficiencies of transfected cells were four times greater than control cells in anchorage-independent assays. Control cells did not form tumors at 5 x 105 in athymic nude mice. Antisense-transfected cells formed tumors in 40{\%} of animals injected. At higher inocula (1 x 106 cells) antisense-transfected cells formed tumors in 100{\%} of animals injected, but control cells still failed to form tumors. These results show that TGF-β1 acts as a negative growth regulator of human colon carcinoma cells in vivo as well as in vitro. Acquisition of partial or full resistance to such inhibitory effects may therefore contribute to tumor development and progression.",
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AU - Willson, J. K.V.

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AU - Humphrey, L. E.

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