Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin

G. M. Kolfschoten, T. M. Hulscher, S. M. Schrier, V. M M Van Houten, H. M. Pinedo, E. Boven

Research output: Contribution to journalArticle

Abstract

Objectives. Apoptosis is believed to be a major mechanism of cisplatin-induced cell death. We investigated the kinetics of apoptosis in four human ovarian cancer cell lines treated with cisplatin to obtain insight into the role and the behavior of a variety of factors involved in this process. Methods. The cell lines A2780, H134, and IGROV-1 (all wild-type p53) and OVCAR-3 (mutant p53) were exposed to cisplatin for 1 h and the antiproliferative effects were measured after 96 h. At various time points up to 96 h after the 1-h exposure to the individual 90% growth-inhibiting cisplatin concentrations, FACS analysis and May-Grünwald Giemsa staining were carried out to determine the extent of apoptosis. At the same time points protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 and the activity of caspase-3 were measured. FACS analysis was also carried out to determine changes in cell cycle distribution as a response to cisplatin. Results. The four cell lines differed in sensitivity to cisplatin. A2780 was the most sensitive and IGROV-1 was the least sensitive. In contrast, IGROV-1 cells showed the highest percentage of apoptosis (30-40%), while A2780 had the lowest percentage (6-14%) (r = 0.99). The occurrence of apoptosis was not dependent on functional p53. Of interest, caspase-3 activity was in line with the percentage of apoptosis and preceded DNA fragmentation and the visualization of condensed nuclei. Wild-type p53 cells accumulated in the S phase, while OVCAR-3 arrested in the G2/M phase. The protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 varied in time, but were not related to the apoptotic behavior of the cells. Upregulation of p53 was already evident before activation of caspase-3. Conclusions. Time-dependent changes in the various factors involved in the apoptotic process induced by equitoxic doses of cisplatin vary strongly among the cell lines. Caspase-3 activation plays an important role in cisplatin-induced apoptosis and this precedes morphological changes. The ability of cells to enter apoptosis, however, does not seem to predict sensitivity to cisplatin.

Original languageEnglish (US)
Pages (from-to)404-412
Number of pages9
JournalGynecologic Oncology
Volume84
Issue number3
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Ovarian Neoplasms
Cisplatin
Apoptosis
Caspase 3
Cell Line
G2 Phase
DNA Fragmentation
S Phase
Cell Division
Cell Cycle
Proteins
Cell Death
Up-Regulation
Staining and Labeling
Growth

Keywords

  • Apoptosis
  • Bax, Bcl-2
  • Caspase-3
  • Cisplatin
  • Human ovarian cancer cell lines
  • p21/WAF1
  • p53

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Oncology

Cite this

Kolfschoten, G. M., Hulscher, T. M., Schrier, S. M., Van Houten, V. M. M., Pinedo, H. M., & Boven, E. (2002). Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin. Gynecologic Oncology, 84(3), 404-412. https://doi.org/10.1006/gyno.2001.6537

Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin. / Kolfschoten, G. M.; Hulscher, T. M.; Schrier, S. M.; Van Houten, V. M M; Pinedo, H. M.; Boven, E.

In: Gynecologic Oncology, Vol. 84, No. 3, 2002, p. 404-412.

Research output: Contribution to journalArticle

Kolfschoten, GM, Hulscher, TM, Schrier, SM, Van Houten, VMM, Pinedo, HM & Boven, E 2002, 'Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin', Gynecologic Oncology, vol. 84, no. 3, pp. 404-412. https://doi.org/10.1006/gyno.2001.6537
Kolfschoten, G. M. ; Hulscher, T. M. ; Schrier, S. M. ; Van Houten, V. M M ; Pinedo, H. M. ; Boven, E. / Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin. In: Gynecologic Oncology. 2002 ; Vol. 84, No. 3. pp. 404-412.
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abstract = "Objectives. Apoptosis is believed to be a major mechanism of cisplatin-induced cell death. We investigated the kinetics of apoptosis in four human ovarian cancer cell lines treated with cisplatin to obtain insight into the role and the behavior of a variety of factors involved in this process. Methods. The cell lines A2780, H134, and IGROV-1 (all wild-type p53) and OVCAR-3 (mutant p53) were exposed to cisplatin for 1 h and the antiproliferative effects were measured after 96 h. At various time points up to 96 h after the 1-h exposure to the individual 90{\%} growth-inhibiting cisplatin concentrations, FACS analysis and May-Gr{\"u}nwald Giemsa staining were carried out to determine the extent of apoptosis. At the same time points protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 and the activity of caspase-3 were measured. FACS analysis was also carried out to determine changes in cell cycle distribution as a response to cisplatin. Results. The four cell lines differed in sensitivity to cisplatin. A2780 was the most sensitive and IGROV-1 was the least sensitive. In contrast, IGROV-1 cells showed the highest percentage of apoptosis (30-40{\%}), while A2780 had the lowest percentage (6-14{\%}) (r = 0.99). The occurrence of apoptosis was not dependent on functional p53. Of interest, caspase-3 activity was in line with the percentage of apoptosis and preceded DNA fragmentation and the visualization of condensed nuclei. Wild-type p53 cells accumulated in the S phase, while OVCAR-3 arrested in the G2/M phase. The protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 varied in time, but were not related to the apoptotic behavior of the cells. Upregulation of p53 was already evident before activation of caspase-3. Conclusions. Time-dependent changes in the various factors involved in the apoptotic process induced by equitoxic doses of cisplatin vary strongly among the cell lines. Caspase-3 activation plays an important role in cisplatin-induced apoptosis and this precedes morphological changes. The ability of cells to enter apoptosis, however, does not seem to predict sensitivity to cisplatin.",
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T1 - Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin

AU - Kolfschoten, G. M.

AU - Hulscher, T. M.

AU - Schrier, S. M.

AU - Van Houten, V. M M

AU - Pinedo, H. M.

AU - Boven, E.

PY - 2002

Y1 - 2002

N2 - Objectives. Apoptosis is believed to be a major mechanism of cisplatin-induced cell death. We investigated the kinetics of apoptosis in four human ovarian cancer cell lines treated with cisplatin to obtain insight into the role and the behavior of a variety of factors involved in this process. Methods. The cell lines A2780, H134, and IGROV-1 (all wild-type p53) and OVCAR-3 (mutant p53) were exposed to cisplatin for 1 h and the antiproliferative effects were measured after 96 h. At various time points up to 96 h after the 1-h exposure to the individual 90% growth-inhibiting cisplatin concentrations, FACS analysis and May-Grünwald Giemsa staining were carried out to determine the extent of apoptosis. At the same time points protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 and the activity of caspase-3 were measured. FACS analysis was also carried out to determine changes in cell cycle distribution as a response to cisplatin. Results. The four cell lines differed in sensitivity to cisplatin. A2780 was the most sensitive and IGROV-1 was the least sensitive. In contrast, IGROV-1 cells showed the highest percentage of apoptosis (30-40%), while A2780 had the lowest percentage (6-14%) (r = 0.99). The occurrence of apoptosis was not dependent on functional p53. Of interest, caspase-3 activity was in line with the percentage of apoptosis and preceded DNA fragmentation and the visualization of condensed nuclei. Wild-type p53 cells accumulated in the S phase, while OVCAR-3 arrested in the G2/M phase. The protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 varied in time, but were not related to the apoptotic behavior of the cells. Upregulation of p53 was already evident before activation of caspase-3. Conclusions. Time-dependent changes in the various factors involved in the apoptotic process induced by equitoxic doses of cisplatin vary strongly among the cell lines. Caspase-3 activation plays an important role in cisplatin-induced apoptosis and this precedes morphological changes. The ability of cells to enter apoptosis, however, does not seem to predict sensitivity to cisplatin.

AB - Objectives. Apoptosis is believed to be a major mechanism of cisplatin-induced cell death. We investigated the kinetics of apoptosis in four human ovarian cancer cell lines treated with cisplatin to obtain insight into the role and the behavior of a variety of factors involved in this process. Methods. The cell lines A2780, H134, and IGROV-1 (all wild-type p53) and OVCAR-3 (mutant p53) were exposed to cisplatin for 1 h and the antiproliferative effects were measured after 96 h. At various time points up to 96 h after the 1-h exposure to the individual 90% growth-inhibiting cisplatin concentrations, FACS analysis and May-Grünwald Giemsa staining were carried out to determine the extent of apoptosis. At the same time points protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 and the activity of caspase-3 were measured. FACS analysis was also carried out to determine changes in cell cycle distribution as a response to cisplatin. Results. The four cell lines differed in sensitivity to cisplatin. A2780 was the most sensitive and IGROV-1 was the least sensitive. In contrast, IGROV-1 cells showed the highest percentage of apoptosis (30-40%), while A2780 had the lowest percentage (6-14%) (r = 0.99). The occurrence of apoptosis was not dependent on functional p53. Of interest, caspase-3 activity was in line with the percentage of apoptosis and preceded DNA fragmentation and the visualization of condensed nuclei. Wild-type p53 cells accumulated in the S phase, while OVCAR-3 arrested in the G2/M phase. The protein expression levels of p53, p21/WAF1, Bax, and Bcl-2 varied in time, but were not related to the apoptotic behavior of the cells. Upregulation of p53 was already evident before activation of caspase-3. Conclusions. Time-dependent changes in the various factors involved in the apoptotic process induced by equitoxic doses of cisplatin vary strongly among the cell lines. Caspase-3 activation plays an important role in cisplatin-induced apoptosis and this precedes morphological changes. The ability of cells to enter apoptosis, however, does not seem to predict sensitivity to cisplatin.

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