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. 2002 Jan 7;86(1):143–150. doi: 10.1038/sj.bjc.6600025

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Copyright © 2002 The Cancer Research Campaign

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Effects of vinflunine on caspases 3/7 activity and PARP cleavage. (A) Enzymatic Ac-DEVD-p-NA cleavage activity in P388 leukaemia cell extracts following a 24-h incubation with vinflunine. Twenty μg protein from cell extracts prepared from cells exposed to 0 (▴), 0.05 (▪), 0.15 (⧫), 0.3 (•) μM vinflunine for 24 h were assayed for caspases-3/7 dependent activity using the synthetic substrate Ac-DEVD-p-NA. Caspases-3/7 activity was also evaluated in presence of 200 μM Ac-DEVD-CHO, the caspases-3/7 inhibitor (μ), for the cell extracts treated with 0.3 μM vinflunine. Assays were performed in triplicates and results were expressed as average optical density values±s.e.m. (B) Cleavage of PARP after either a 24 h-incubation of P388 cells with 0.1–0.5 μM vinflunine (B1) or a 3–24-h incubation with 0.3 μM vinflunine (B2). Vinflunine-induced PARP cleavage was analyzed by Western blot. Results are expressed as means from 2–3 independent experiments of the percentage of degraded form vs total (degraded plus native) PARP forms±s.e.m.