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Dynamics of cell cycle phase perturbations by trabectedin (ET-743) in nucleotide excision repair (NER)-deficient and NER-proficient cells, unravelled by a novel mathematical simulation approach

Tavecchio, M LU ; Natoli, C; Ubezio, P; Erba, Eugenio and D'Incalci, Maurizio (2007) In Cell Proliferation 40(6). p.885-904
Abstract

OBJECTIVES: Trabectedin (ET-743, Yondelis) is a natural marine product, with antitumour activity, currently in phase II/III clinical trials. Previous studies have shown that cells hypersensitive to ultraviolet (UV)-rays because of nucleotide excision repair (NER) deficiency, were resistant to trabectedin. The purpose of this study was to investigate whether this resistance was associated with different drug-induced cell cycle perturbations.

MATERIALS AND METHODS: An isogenic NER-proficient cellular system (CHO-AA8) and a NER-deficient one (CHO-UV-96), lacking functional ERCC-1, were studied. Flow cytometric assays showed progressive accumulation of cells in G2 + M phase in NER-proficient but not in NER-deficient cells. Applying a... (More)

OBJECTIVES: Trabectedin (ET-743, Yondelis) is a natural marine product, with antitumour activity, currently in phase II/III clinical trials. Previous studies have shown that cells hypersensitive to ultraviolet (UV)-rays because of nucleotide excision repair (NER) deficiency, were resistant to trabectedin. The purpose of this study was to investigate whether this resistance was associated with different drug-induced cell cycle perturbations.

MATERIALS AND METHODS: An isogenic NER-proficient cellular system (CHO-AA8) and a NER-deficient one (CHO-UV-96), lacking functional ERCC-1, were studied. Flow cytometric assays showed progressive accumulation of cells in G2 + M phase in NER-proficient but not in NER-deficient cells. Applying a computer simulation method, we realized that the dynamics of the cell cycle perturbations in all phases were complex.

RESULTS: Cells exposed to trabectedin during G1 and G2 + M first experienced a G1 block, while those exposed in S phase were delayed in S and G2 + M phases but eventually divided. In the presence of functional NER, exit from the G1 block was faster; then, cells progressed slowly through S phase and were subsequently blocked in G2 + M phase. This G2 + M processing of trabectedin-induced damage in NER-proficient cells was unable to restore cell cycling, suggesting a difficulty in repairing the damage.

CONCLUSIONS: This might be due either to important damage left unrepaired by previous G1 repair, or that NER activity itself caused DNA damage, or both. We speculate that in UV-96 cells repair mechanisms other than NER are activated both in G1 and G2 + M phases.

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publication status
published
keywords
Animals, Bromodeoxyuridine, CHO Cells, Cell Count, Cell Cycle, Cell Death, Cell Line, Cell Proliferation, Colony-Forming Units Assay, Computer Simulation, Cricetinae, Cricetulus, DNA, DNA Repair, Dioxoles, Flow Cytometry, G1 Phase, G2 Phase, Models, Biological, S Phase, Tetrahydroisoquinolines, Journal Article, Research Support, Non-U.S. Gov't
in
Cell Proliferation
volume
40
issue
6
pages
885 - 904
publisher
Wiley-Blackwell
external identifiers
  • scopus:36248974108
ISSN
1365-2184
DOI
10.1111/j.1365-2184.2007.00469.x
language
English
LU publication?
no
id
21cb4737-f531-4250-a70e-909198eb5aa6
date added to LUP
2017-03-07 09:15:06
date last changed
2017-11-12 04:30:15
@article{21cb4737-f531-4250-a70e-909198eb5aa6,
  abstract     = {<p>OBJECTIVES: Trabectedin (ET-743, Yondelis) is a natural marine product, with antitumour activity, currently in phase II/III clinical trials. Previous studies have shown that cells hypersensitive to ultraviolet (UV)-rays because of nucleotide excision repair (NER) deficiency, were resistant to trabectedin. The purpose of this study was to investigate whether this resistance was associated with different drug-induced cell cycle perturbations.</p><p>MATERIALS AND METHODS: An isogenic NER-proficient cellular system (CHO-AA8) and a NER-deficient one (CHO-UV-96), lacking functional ERCC-1, were studied. Flow cytometric assays showed progressive accumulation of cells in G2 + M phase in NER-proficient but not in NER-deficient cells. Applying a computer simulation method, we realized that the dynamics of the cell cycle perturbations in all phases were complex.</p><p>RESULTS: Cells exposed to trabectedin during G1 and G2 + M first experienced a G1 block, while those exposed in S phase were delayed in S and G2 + M phases but eventually divided. In the presence of functional NER, exit from the G1 block was faster; then, cells progressed slowly through S phase and were subsequently blocked in G2 + M phase. This G2 + M processing of trabectedin-induced damage in NER-proficient cells was unable to restore cell cycling, suggesting a difficulty in repairing the damage.</p><p>CONCLUSIONS: This might be due either to important damage left unrepaired by previous G1 repair, or that NER activity itself caused DNA damage, or both. We speculate that in UV-96 cells repair mechanisms other than NER are activated both in G1 and G2 + M phases.</p>},
  author       = {Tavecchio, M and Natoli, C and Ubezio, P and Erba, Eugenio and D'Incalci, Maurizio},
  issn         = {1365-2184},
  keyword      = {Animals,Bromodeoxyuridine,CHO Cells,Cell Count,Cell Cycle,Cell Death,Cell Line,Cell Proliferation,Colony-Forming Units Assay,Computer Simulation,Cricetinae,Cricetulus,DNA,DNA Repair,Dioxoles,Flow Cytometry,G1 Phase,G2 Phase,Models, Biological,S Phase,Tetrahydroisoquinolines,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  number       = {6},
  pages        = {885--904},
  publisher    = {Wiley-Blackwell},
  series       = {Cell Proliferation},
  title        = {Dynamics of cell cycle phase perturbations by trabectedin (ET-743) in nucleotide excision repair (NER)-deficient and NER-proficient cells, unravelled by a novel mathematical simulation approach},
  url          = {http://dx.doi.org/10.1111/j.1365-2184.2007.00469.x},
  volume       = {40},
  year         = {2007},
}