DNA damage cell checkpoint activities are altered in monocrotaline pyrrole-induced cell cycle arrest in human pulmonary artery endothelial cells

Toxicol Appl Pharmacol. 2000 Jul 15;166(2):69-80. doi: 10.1006/taap.2000.8966.


Monocrotaline pyrrole (MCTP) causes cyto- and karyomegaly and persistent cell cycle arrest in the G2 stage of the cell cycle in cultured bovine pulmonary artery endothelial cells. To better characterize the cell cycle regulatory mechanisms of this process as well as determine whether this process would occur in cells of human origin, we treated human pulmonary artery endothelial cell (HPAEC) cultures with MCTP and determined, by flow cytometry, the expression of cyclin B1 and p53 in conjunction with DNA content. We also validated by Western blots that the persistence of cdc2 in its inactivated phosphorylated state, previously described in bovine cell cultures, occurred in HPAEC. Alterations in p53, cyclin A, cyclin B1, and cdc25c expression were also examined in Western blots of treated HPAEC extracts. The response of HPAEC to MCTP was compared with that of adriamycin and nocodazole, agents known to cause cell cycle alterations. Results of these experiments demonstrate that HPAEC treated with MCTP develop a population of cells in G2 that has increased cyclin B1 expression. These cells express increased amounts of cdc2 but not cdc25c. The ratio of inactive triphosphorylated cdc2 to the active monophosphorylated form increased moderately from control cultures in contrast to predominance of the active form in nocodazole-treated cultures. In addition, a second population of cells expressing cyclin B1 had continued incorporation of BrdU and DNA content consistent with 8 N chromosomes. A similar 8 N cell population was evident in nocodazole-treated cells but these cells had both cyclin B1 positive and negative components. Compared with adriamycin, a known inducer of p53, MCTP-treated HPAEC expressed p53 only at high concentrations and p53 expression was not coordinated with G2 arrest or polyploidy. We conclude that HPAEC treated with low concentrations of MCTP develop G2 arrest in association with persistent cyclin B1 expression, failure to completely activate cdc2, and continued DNA synthesis through a pathway that is unrelated to altered expression of p53.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Animals
  • Blotting, Western
  • Cattle
  • Cell Cycle / drug effects*
  • Cell Cycle Proteins / metabolism
  • Cells, Cultured
  • Child
  • Cyclin A / metabolism
  • Cyclin B / metabolism
  • Cyclin B1
  • DNA / analysis
  • DNA Damage / drug effects*
  • Doxorubicin / pharmacology
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / metabolism
  • Female
  • Flow Cytometry
  • G2 Phase / drug effects
  • G2 Phase / physiology
  • Humans
  • Male
  • Monocrotaline / analogs & derivatives*
  • Monocrotaline / toxicity
  • Nocodazole / pharmacology
  • Pulmonary Artery / drug effects*
  • Pulmonary Artery / metabolism
  • Tumor Suppressor Protein p53 / metabolism
  • cdc25 Phosphatases / metabolism


  • CCNB1 protein, human
  • Cell Cycle Proteins
  • Cyclin A
  • Cyclin B
  • Cyclin B1
  • Tumor Suppressor Protein p53
  • monocrotaline pyrrole
  • Monocrotaline
  • Doxorubicin
  • DNA
  • CDC25C protein, human
  • cdc25 Phosphatases
  • Nocodazole