Transforming growth factor-beta1 inhibits generation of angiostatin by human pancreatic cancer cells

C. A. O'Mahony, Daniel Albo, G. P. Tuszynski, D. H. Berger, R. D. Beauchamp, T. C. Ko

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Background. Angiostatin, a proteolytic fragment of plasminogen, is a potent inhibitor of angiogenesis. We have previously shown that the human pancreatic cancer cell line ASPC-1 produces enzymatic activity capable of generating angiostatin. In this study we sought to determine whether angiostatin production by ASPC-1 cells was regulated by the growth factor transforming growth factor-β1 (TGF-β1), a key mediator of tumor angiogenesis. Methods. ASPC-1 cells were grown to 70% to 80% confluence in 20% fetal calf serum-RPMI. Medium was changed to serum free. TGF-β1 was added at concentrations of 0, 1, 5, and 10 ng/mL with or without plasminogen activator inhibitor type-1 (PAI-1) at concentrations of 0, 5, 10, 50, and 100 μg/mL. Cells were then cultured for an additional 24 hours. The serum-free conditioned medium was obtained. Angiostatin generation was determined by incubating 20 μg of plasminogen with 100 IlL of serum-free conditioned medium for 0, 1, 2, 3, 6, 12, and 24 hours. Samples were run on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred. The membrane was probed with a monoclonal antibody to the kringle 1-3 fragment of plasminogen and developed using enhanced chemiluminesence. Results. TGF-β1 and PAI-1 inhibited the conversion of plasminogen into angiostatin in a time- and dose-dependent manner. Antibody to PAId completely blocks TGF-β1 mediated angiostatin inhibition. Conclusions. TGF-β1 inhibits the generation of the antiangiogenic molecule angiostatin by human pancreatic cancer cells in a time- and dose-dependent manner. This effect is mediated through modulation of the plasminogen/plasmin system.

Original languageEnglish (US)
Pages (from-to)388-393
Number of pages6
JournalSurgery
Volume124
Issue number2
DOIs
StatePublished - Jan 1 1998
Externally publishedYes

Fingerprint

Angiostatins
Transforming Growth Factor beta1
Pancreatic Neoplasms
Plasminogen
Transforming Growth Factors
Plasminogen Activator Inhibitor 1
Serum-Free Culture Media
Conditioned Culture Medium
Kringles
Angiogenesis Inhibitors
Fibrinolysin
Serum
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Intercellular Signaling Peptides and Proteins
Monoclonal Antibodies
Cell Line
Membranes
Antibodies

ASJC Scopus subject areas

  • Surgery

Cite this

O'Mahony, C. A., Albo, D., Tuszynski, G. P., Berger, D. H., Beauchamp, R. D., & Ko, T. C. (1998). Transforming growth factor-beta1 inhibits generation of angiostatin by human pancreatic cancer cells. Surgery, 124(2), 388-393. https://doi.org/10.1016/S0039-6060(98)70145-X

Transforming growth factor-beta1 inhibits generation of angiostatin by human pancreatic cancer cells. / O'Mahony, C. A.; Albo, Daniel; Tuszynski, G. P.; Berger, D. H.; Beauchamp, R. D.; Ko, T. C.

In: Surgery, Vol. 124, No. 2, 01.01.1998, p. 388-393.

Research output: Contribution to journalArticle

O'Mahony, C. A. ; Albo, Daniel ; Tuszynski, G. P. ; Berger, D. H. ; Beauchamp, R. D. ; Ko, T. C. / Transforming growth factor-beta1 inhibits generation of angiostatin by human pancreatic cancer cells. In: Surgery. 1998 ; Vol. 124, No. 2. pp. 388-393.
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abstract = "Background. Angiostatin, a proteolytic fragment of plasminogen, is a potent inhibitor of angiogenesis. We have previously shown that the human pancreatic cancer cell line ASPC-1 produces enzymatic activity capable of generating angiostatin. In this study we sought to determine whether angiostatin production by ASPC-1 cells was regulated by the growth factor transforming growth factor-β1 (TGF-β1), a key mediator of tumor angiogenesis. Methods. ASPC-1 cells were grown to 70{\%} to 80{\%} confluence in 20{\%} fetal calf serum-RPMI. Medium was changed to serum free. TGF-β1 was added at concentrations of 0, 1, 5, and 10 ng/mL with or without plasminogen activator inhibitor type-1 (PAI-1) at concentrations of 0, 5, 10, 50, and 100 μg/mL. Cells were then cultured for an additional 24 hours. The serum-free conditioned medium was obtained. Angiostatin generation was determined by incubating 20 μg of plasminogen with 100 IlL of serum-free conditioned medium for 0, 1, 2, 3, 6, 12, and 24 hours. Samples were run on 12{\%} sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred. The membrane was probed with a monoclonal antibody to the kringle 1-3 fragment of plasminogen and developed using enhanced chemiluminesence. Results. TGF-β1 and PAI-1 inhibited the conversion of plasminogen into angiostatin in a time- and dose-dependent manner. Antibody to PAId completely blocks TGF-β1 mediated angiostatin inhibition. Conclusions. TGF-β1 inhibits the generation of the antiangiogenic molecule angiostatin by human pancreatic cancer cells in a time- and dose-dependent manner. This effect is mediated through modulation of the plasminogen/plasmin system.",
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N2 - Background. Angiostatin, a proteolytic fragment of plasminogen, is a potent inhibitor of angiogenesis. We have previously shown that the human pancreatic cancer cell line ASPC-1 produces enzymatic activity capable of generating angiostatin. In this study we sought to determine whether angiostatin production by ASPC-1 cells was regulated by the growth factor transforming growth factor-β1 (TGF-β1), a key mediator of tumor angiogenesis. Methods. ASPC-1 cells were grown to 70% to 80% confluence in 20% fetal calf serum-RPMI. Medium was changed to serum free. TGF-β1 was added at concentrations of 0, 1, 5, and 10 ng/mL with or without plasminogen activator inhibitor type-1 (PAI-1) at concentrations of 0, 5, 10, 50, and 100 μg/mL. Cells were then cultured for an additional 24 hours. The serum-free conditioned medium was obtained. Angiostatin generation was determined by incubating 20 μg of plasminogen with 100 IlL of serum-free conditioned medium for 0, 1, 2, 3, 6, 12, and 24 hours. Samples were run on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred. The membrane was probed with a monoclonal antibody to the kringle 1-3 fragment of plasminogen and developed using enhanced chemiluminesence. Results. TGF-β1 and PAI-1 inhibited the conversion of plasminogen into angiostatin in a time- and dose-dependent manner. Antibody to PAId completely blocks TGF-β1 mediated angiostatin inhibition. Conclusions. TGF-β1 inhibits the generation of the antiangiogenic molecule angiostatin by human pancreatic cancer cells in a time- and dose-dependent manner. This effect is mediated through modulation of the plasminogen/plasmin system.

AB - Background. Angiostatin, a proteolytic fragment of plasminogen, is a potent inhibitor of angiogenesis. We have previously shown that the human pancreatic cancer cell line ASPC-1 produces enzymatic activity capable of generating angiostatin. In this study we sought to determine whether angiostatin production by ASPC-1 cells was regulated by the growth factor transforming growth factor-β1 (TGF-β1), a key mediator of tumor angiogenesis. Methods. ASPC-1 cells were grown to 70% to 80% confluence in 20% fetal calf serum-RPMI. Medium was changed to serum free. TGF-β1 was added at concentrations of 0, 1, 5, and 10 ng/mL with or without plasminogen activator inhibitor type-1 (PAI-1) at concentrations of 0, 5, 10, 50, and 100 μg/mL. Cells were then cultured for an additional 24 hours. The serum-free conditioned medium was obtained. Angiostatin generation was determined by incubating 20 μg of plasminogen with 100 IlL of serum-free conditioned medium for 0, 1, 2, 3, 6, 12, and 24 hours. Samples were run on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred. The membrane was probed with a monoclonal antibody to the kringle 1-3 fragment of plasminogen and developed using enhanced chemiluminesence. Results. TGF-β1 and PAI-1 inhibited the conversion of plasminogen into angiostatin in a time- and dose-dependent manner. Antibody to PAId completely blocks TGF-β1 mediated angiostatin inhibition. Conclusions. TGF-β1 inhibits the generation of the antiangiogenic molecule angiostatin by human pancreatic cancer cells in a time- and dose-dependent manner. This effect is mediated through modulation of the plasminogen/plasmin system.

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