Guanylyl cyclase C agonists regulate progression through the cell cycle of human colon carcinoma cells

G. M. Pitari, M. D. Di Guglielmo, J. Park, S. Schulz, S. A. Waldman

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

The effects of Escherichia coli heat-stable enterotoxin (ST) and uroguanylin were examined on the proliferation of T84 and Caco2 human colon carcinoma cells that express guanylyl cyclase C (GC-C) and SW480 human colon carcinoma cells that do not express this receptor. ST or uroguanylin inhibited proliferation of T84 and Caco2 cells, but not SW480 cells, in a concentration-dependent fashion, assessed by quantifying cell number, cell protein, and [3H]thymidine incorporation into DNA. These agonists did not inhibit proliferation by induction of apoptosis, assessed by TUNEL (terminal deoxynucleotidyl transferase-mediated dNTP-biotin nick end labeling of DNA fragments) assay and DNA laddering, or necrosis, assessed by trypan blue exclusion and lactate dehydrogenase release. Rather, ST prolonged the cell cycle, assessed by flow cytometry and [3H]thymidine incorporation into DNA. The cytostatic effects of GC-C agonists were associated with accumulation of intracellular cGMP, mimicked by the cell-permeant analog 8-Br-cGMP, and reproduced and potentiated by the cGMP-specific phosphodiesterase inhibitor zaprinast but not the inactive ST analog TJU 1-103. Thus, GC-C agonists regulate the proliferation of intestinal cells through cGMP-dependent mechanisms by delaying progression of the cell cycle. These data suggest that endogenous agonists of GC-C, such as uroguanylin, may play a role in regulating the balance between epithelial proliferation and differentiation in normal intestinal physiology. Therefore, GC-C ligands may be novel therapeutic agents for the treatment of patients with colorectal cancer.

Original languageEnglish (US)
Pages (from-to)7846-7851
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number14
DOIs
StatePublished - 2001

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Cell Cycle
Colon
Carcinoma
DNA
Thymidine
Phosphodiesterase Inhibitors
Trypan Blue
DNA Nucleotidylexotransferase
In Situ Nick-End Labeling
Cytostatic Agents
Proxy
Biotin
L-Lactate Dehydrogenase
enterotoxin receptor
Colorectal Neoplasms
Flow Cytometry
Necrosis
Cell Count
Hot Temperature
Cell Proliferation

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Guanylyl cyclase C agonists regulate progression through the cell cycle of human colon carcinoma cells. / Pitari, G. M.; Di Guglielmo, M. D.; Park, J.; Schulz, S.; Waldman, S. A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 14, 2001, p. 7846-7851.

Research output: Contribution to journalArticle

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abstract = "The effects of Escherichia coli heat-stable enterotoxin (ST) and uroguanylin were examined on the proliferation of T84 and Caco2 human colon carcinoma cells that express guanylyl cyclase C (GC-C) and SW480 human colon carcinoma cells that do not express this receptor. ST or uroguanylin inhibited proliferation of T84 and Caco2 cells, but not SW480 cells, in a concentration-dependent fashion, assessed by quantifying cell number, cell protein, and [3H]thymidine incorporation into DNA. These agonists did not inhibit proliferation by induction of apoptosis, assessed by TUNEL (terminal deoxynucleotidyl transferase-mediated dNTP-biotin nick end labeling of DNA fragments) assay and DNA laddering, or necrosis, assessed by trypan blue exclusion and lactate dehydrogenase release. Rather, ST prolonged the cell cycle, assessed by flow cytometry and [3H]thymidine incorporation into DNA. The cytostatic effects of GC-C agonists were associated with accumulation of intracellular cGMP, mimicked by the cell-permeant analog 8-Br-cGMP, and reproduced and potentiated by the cGMP-specific phosphodiesterase inhibitor zaprinast but not the inactive ST analog TJU 1-103. Thus, GC-C agonists regulate the proliferation of intestinal cells through cGMP-dependent mechanisms by delaying progression of the cell cycle. These data suggest that endogenous agonists of GC-C, such as uroguanylin, may play a role in regulating the balance between epithelial proliferation and differentiation in normal intestinal physiology. Therefore, GC-C ligands may be novel therapeutic agents for the treatment of patients with colorectal cancer.",
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