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Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin.
Gynecol Oncol. 2002 Mar; 84(3):404-12.GO

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.

Authors+Show Affiliations

Kolfschoten GM
Department of Medical Oncology, Vrije Universiteit Medical Centre, Amsterdam, the Netherlands.
Hulscher TM
No affiliation info available
Schrier SM
No affiliation info available
van Houten VM
No affiliation info available
Pinedo HM
No affiliation info available
Boven E
No affiliation info available

MeSH

Antineoplastic AgentsApoptosisCaspase 3CaspasesCell CycleCisplatinCyclin-Dependent Kinase Inhibitor p21CyclinsEnzyme ActivationFemaleHumansInhibitory Concentration 50Ovarian NeoplasmsProto-Oncogene ProteinsProto-Oncogene Proteins c-bcl-2Tumor Cells, CulturedTumor Suppressor Protein p53bcl-2-Associated X Protein

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11855878

Citation

Kolfschoten, G M., et al. "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, 2002, pp. 404-12.
Kolfschoten GM, Hulscher TM, Schrier SM, et al. Time-dependent changes in factors involved in the apoptotic process in human ovarian cancer cells as a response to cisplatin. Gynecol Oncol. 2002;84(3):404-12.
Kolfschoten, G. M., Hulscher, T. M., Schrier, S. M., van Houten, V. 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), 404-12.
Kolfschoten GM, et al. Time-dependent Changes in Factors Involved in the Apoptotic Process in Human Ovarian Cancer Cells as a Response to Cisplatin. Gynecol Oncol. 2002;84(3):404-12. PubMed PMID: 11855878.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR 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 - 2002/2/22/pubmed PY - 2002/3/20/medline PY - 2002/2/22/entrez SP - 404 EP - 12 JF - Gynecologic oncology JO - Gynecol Oncol VL - 84 IS - 3 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. SN - 0090-8258 UR - https://www.unboundmedicine.com/medline/citation/11855878/Time_dependent_changes_in_factors_involved_in_the_apoptotic_process_in_human_ovarian_cancer_cells_as_a_response_to_cisplatin_ DB - PRIME DP - Unbound Medicine ER -