Prolonged cell-cycle arrest associated with altered cdc2 kinase in monocrotaline pyrrole-treated pulmonary artery endothelial cells

Heath C. Thomas, Michael W. Lamé, Dexter Morin, Dennis W Wilson, H. J. Segall

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21 Citations (Scopus)

Abstract

Monocrotaline pyrrole (MCTP), a metabolite of the pyrrolizidine alkaloid monocrotaline, is thought to initiate damage to pulmonary endothelial cells resulting in delayed but progressive pulmonary interstitial edema, vascular wall remodeling, and increasing pulmonary hypertension. MCTP was previously shown to inhibit pulmonary endothelial cell proliferation and cause cell-cycle arrest in vitro. To determine the persistence of arrest and better characterize the cell-cycle stage at which it occurs, bovine pulmonary artery endothelial cells (BPAEC) under differing growth conditions were exposed to low (5 μg/ml) or high (34.5 μg/ml) concentrations of MCTP for varying times. Flow cytometric cell-cycle analysis was coupled with Western blot and biochemical analysis of cdc2 kinase and measurements of cell size. MCTP treatment induced a G2 + M phase arrest in 48-h exposed confluent BPAEC that persisted for at least 28 d and was associated with continued cellular enlargement. A short-duration MCTP exposure of confluent (low and high concentration) and log phase (high concentration) BPAEC caused persistent cell-cycle arrest for 1 wk, whereas a low-concentration exposure in log phase cells resulted in cell-cycle arrest with reversal 96 h after exposure. Western blot examination revealed that by 24 h of MCTP exposure, the phosphorylation state of cdc2 was consistent with the inactive form of the kinase (confirmed by biochemical assay); this alteration persisted through at least 96 h of exposure. We conclude that MCTP induces a progressive irreversible endothelial cell dysfunction leading to inactivation of cdc2 kinase and irreversible cell-cycle arrest at the G2 checkpoint.

Original languageEnglish (US)
Pages (from-to)129-142
Number of pages14
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume19
Issue number1
StatePublished - 1998

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Endothelial cells
Cell Cycle Checkpoints
Pulmonary Artery
Phosphotransferases
Endothelial Cells
Cells
Cell Cycle
Western Blotting
Monocrotaline
G2 Phase Cell Cycle Checkpoints
Pyrrolizidine Alkaloids
Lung
Phosphorylation
G2 Phase
Cell proliferation
Pulmonary Edema
Metabolites
monocrotaline pyrrole
Cell Size
Pulmonary Hypertension

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

Cite this

Prolonged cell-cycle arrest associated with altered cdc2 kinase in monocrotaline pyrrole-treated pulmonary artery endothelial cells. / Thomas, Heath C.; Lamé, Michael W.; Morin, Dexter; Wilson, Dennis W; Segall, H. J.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 19, No. 1, 1998, p. 129-142.

Research output: Contribution to journalArticle

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