Cisplatin Induces Apoptosis in a Human Ovarian Carcinoma Cell Line without Concomitant Internucleosomal Degradation of DNA

Ormerod, M.G.; O’Neill, Ciaran; Robertson, David; Harrap, K. R.

doi:10.1006/excr.1994.1082

“…Cisplatin, a widely known DNA alkylating drug and inducer of DSBs, 23 was used to verify whether the findings described above, applied to other DNA-damaging drugs. As shown in Relationship between stress level and incubation time required for the induction of senescence DSBs, and apoptosis…”

Section: Validation For Other Drugs and Cell Typesmentioning

confidence: 99%

The role of histone acetylation versus DNA damage in drug-induced senescence and apoptosis

Rebbaa

¹

,

Zheng

²

,

Chu

³

et al. 2006

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The present study was undertaken to determine the significance of histone acetylation versus DNA damage in drug-induced irreversible growth arrest (senescence) and apoptosis. Cellular treatment with the DNA-damaging drugs doxorubicin and cisplatin or with the histone deacetylase inhibitor trichostatin A, led to the finding that all the three drugs induced senescence at concentrations significantly lower than those required for apoptosis. However, only doxorubicin and cisplatin induced activation of H2AX, a marker for double-strand break formation. Interestingly, this occurred mainly at apoptosis and not senescence-inducing drug concentrations, suggesting that non-DNA-damage pathways may be implicated in induction of senescence by these drugs. In agreement with this, chromatin immunoprecipitation experiments indicated that doxorubicin was able to induce acetylation of histone H3 at the promoter of p21/WAF1 only at senescence-inducing concentrations. Collectively, these findings suggest that alteration of chromatin structure by cytotoxic drugs may represent a key mediator of senescence.

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“…These cells exhibited apoptotic morphology with compacted and fragmented nuclei when compared with attached control cells where the chromatin was diffuse ( Figure 5). Previous studies have shown that the morphology of attached treated cells is similar to that of attached control untreated cells (Ormerod et al, 1994aO'Neill et al, 1996).…”

Section: Morphologymentioning

confidence: 79%

“…However, there was evidence of fragmentation in the attached cell population of both CH1 cell lines running between 90 and 150 kb, which was absent from the SKOV-3 cell line. Earlier studies demonstrated that the DNA from attached control untreated cells does not migrate much beyond the loading point (Ormerod et al, 1994a Cell cycle analysis Flow cytometric analysis was carried out from attached cells collected at various time points after incubation with 2 × IC 50 JM216 (Figure 7). The main cell cycle effect of JM216 was a slow down of traverse through S phase, which culminated in a G2 block occurring from 48 h to 72 h in the CH1 and CH1cisR cell lines following removal of drug.…”

Section: Field Inversion Gel Electrophoresismentioning

confidence: 99%

Gene-specific repair of Pt/DNA lesions and induction of apoptosis by the oral platinum drug JM216 in three human ovarian carcinoma cell lines sensitive and resistant to cisplatin

O’Neill¹,

Koberle²,

Kèlland³

1999

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Summary JM216, an oral platinum drug entering into phase III clinical trial, exhibited comparable cytotoxicity to cisplatin in three human ovarian carcinoma cell lines: the sensitive (CH1), acquired resistant (CH1cisR) and intrinsically resistant (SKOV-3). Platinum accumulation and binding to DNA were similar in each of the three cell lines at equimolar doses, indicating that the resistant cell lines could tolerate higher intracellular platinum levels and platinum bound to DNA at IC 50 concentrations of drug. Comparison with cisplatin demonstrated that intracellular platinum levels were marginally higher with JM216, but that platinum binding to DNA was similar for the two drugs in each of the cell lines. Each of the cell lines exhibited an ability to repair JM216 induced platinum/DNA lesions in the N-ras gene (gene-specific repair) at equitoxic concentrations of drug. However, this occurred to a greater extent in the two resistant cell lines such that by 24 h the CH1cisR and SKOV-3 had removed 72% and 67% respectively compared with approximately 32% for the CH1. Reduced gene-specific repair capacity in CH1 cells was also seen following incubation with 25 µM (or 5 µM -2 × IC 50 ) cisplatin, whereas the CH1cisR and SKOV-3 cell lines were repair proficient. JM216 induced apoptosis in the three cell lines following a 2h incubation with 2 × the IC 50 of drug. Fluorescent microscopy of cells stained with propidium iodide showed that the detached cell population displayed typical apoptotic nuclei. Furthermore, field inversion gel electrophoresis demonstrated the presence of DNA fragments approximately 23-50 kb in size, indicative of apoptosis, in the detached cells. JM216 induced an S phase slow down in each of the three cell lines accompanied by a G2 block in the CH1 pair. Incubation with this concentration of JM216 also resulted in the induction of p53 in the CH1 and CH1cisR. These studies suggest that the relative sensitivity of the CH1 cell line to cisplatin and JM216 is at least partly attributable to a deficiency in gene-specific repair. The oral platinum drug, JM216, exerts its cytotoxic effects through the induction of apoptosis following a slow-down in S phase in both the sensitive and resistant lines.

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“…The difference in the rate of cell death may have been due to the difference in doubling times for the CH1 pair (17 h) and the SKOV-3 (22 h). Apoptotic nuclei were detected by fluorescence microscopy following incubation with PI and the characteristic 23-50 kb DNA fragment associated with apoptosis was detected by FIGE (Oberhammer et al, 1993;Ormerod et al, 1994aOrmerod et al, , 1996O'Neill et al, 1996).…”

Section: Discussionmentioning

confidence: 99%

Gene-specific repair of Pt/DNA lesions and induction of apoptosis by the oral platinum drug JM216 in three human ovarian carcinoma cell lines sensitive and resistant to cisplatin

O’Neill

¹

,

Köberle

²

,

Masters

³

et al. 1999

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Summary JM216, an oral platinum drug entering into phase III clinical trial, exhibited comparable cytotoxicity to cisplatin in three human ovarian carcinoma cell lines: the sensitive (CH1), acquired resistant (CH1cisR) and intrinsically resistant (SKOV-3). Platinum accumulation and binding to DNA were similar in each of the three cell lines at equimolar doses, indicating that the resistant cell lines could tolerate higher intracellular platinum levels and platinum bound to DNA at IC 50 concentrations of drug. Comparison with cisplatin demonstrated that intracellular platinum levels were marginally higher with JM216, but that platinum binding to DNA was similar for the two drugs in each of the cell lines. Each of the cell lines exhibited an ability to repair JM216 induced platinum/DNA lesions in the N-ras gene (gene-specific repair) at equitoxic concentrations of drug. However, this occurred to a greater extent in the two resistant cell lines such that by 24 h the CH1cisR and SKOV-3 had removed 72% and 67% respectively compared with approximately 32% for the CH1. Reduced gene-specific repair capacity in CH1 cells was also seen following incubation with 25 µM (or 5 µM -2 × IC 50 ) cisplatin, whereas the CH1cisR and SKOV-3 cell lines were repair proficient. JM216 induced apoptosis in the three cell lines following a 2h incubation with 2 × the IC 50 of drug. Fluorescent microscopy of cells stained with propidium iodide showed that the detached cell population displayed typical apoptotic nuclei. Furthermore, field inversion gel electrophoresis demonstrated the presence of DNA fragments approximately 23-50 kb in size, indicative of apoptosis, in the detached cells. JM216 induced an S phase slow down in each of the three cell lines accompanied by a G2 block in the CH1 pair. Incubation with this concentration of JM216 also resulted in the induction of p53 in the CH1 and CH1cisR. These studies suggest that the relative sensitivity of the CH1 cell line to cisplatin and JM216 is at least partly attributable to a deficiency in gene-specific repair. The oral platinum drug, JM216, exerts its cytotoxic effects through the induction of apoptosis following a slow-down in S phase in both the sensitive and resistant lines.

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Cisplatin Induces Apoptosis in a Human Ovarian Carcinoma Cell Line without Concomitant Internucleosomal Degradation of DNA

Cited by 112 publications

References 0 publications

The role of histone acetylation versus DNA damage in drug-induced senescence and apoptosis

The role of histone acetylation versus DNA damage in drug-induced senescence and apoptosis

Gene-specific repair of Pt/DNA lesions and induction of apoptosis by the oral platinum drug JM216 in three human ovarian carcinoma cell lines sensitive and resistant to cisplatin

Gene-specific repair of Pt/DNA lesions and induction of apoptosis by the oral platinum drug JM216 in three human ovarian carcinoma cell lines sensitive and resistant to cisplatin

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