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- W2904708432 abstract "3585 Malignant mesothelioma (MM) is highly resistant to drug therapy. Since many cytotoxic agents exert their effect by induction of apoptosis we investigated the genetic control of key regulators of the mitochondrial apoptotic pathway, the Bcl-2 protein family, during cisplatin induced apoptosis in MM. Gene expression was assayed in four representative human MM cell lines (ONE58, JU77, NO36, LO68) using conventional and real-time RT-PCR. The expression and subcellular localisation of p53 was determined by fluorescence microscopy. All four MM cell lines constitutively expressed Bcl-2, Bcl-xL, Bax, Bak, NOXA and PUMA as demonstrated by RT-PCR. Cells were exposed to cisplatin at concentrations of 0-100μg/ml and apoptosis/cytotoxicity was assessed by MTT and JC-1 mitochondrial depolarization assays. Sensitivity to cisplatin as reflected in the IC50 ranged from 1.13±0.2 μg/ml (JU77) to 3.72±1.3 μg/ml (ONE58). Mitochondrial depolarization indicated an active role for the mitochondria during cisplatin induced apoptosis. Cisplatin induction of gene expression was examined in the JU77 and ONE58 at their respective IC50. There were significant differences in the magnitude and timing of transcriptional responses. Bak was a transcriptional target of cisplatin in both cell lines, although maximal upregulation was seen at 1h in JU77 compared to 6h in ONE58. In JU77 Bcl-xL and NOXA mRNAs were also upregulated at 1 hr, but Bax mRNA was not. In contrast, in ONE58 Bcl-xL was essentially unchanged while both Bax and NOXA were upregulated. In ONE58 basal p53 levels were significantly elevated in comparison to JU77. In response to cisplatin p53 levels increased in JU77 but not ONE58. These findings indicate that pro-apoptotic Bak gene expression was a significant target of cisplatin in both JU77 and ONE58. Changes in anti-apoptotic Bcl-xL expression may play a role in the resistance of JU77 but did not appear to be a factor in ONE58. These results provide insights into apoptotic mechanisms in MM cells after exposure to cytotoxic drugs." @default.
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- W2904708432 date "2007-05-01" @default.
- W2904708432 modified "2023-09-23" @default.
- W2904708432 title "Expression of Bcl-2 family genes during cisplatin induced apoptosis in mesothelioma cells" @default.
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