FEBS Letters | |
Protease inhibitors block apoptosis at intermediate stages: a compared analysis of DNA fragmentation and apoptotic nuclear morphology | |
Maresca, V.2  Coppola, S.2  Ghibelli, L.2  Gualandi, G.1  | |
[1] DABAC, Università della Tuscia, 01100 Viterbo, Italy;Dipartimento di Biologia, Università di Roma Tor Vergata, via della Ricerca Scientifica, 00123 Roma, Italy | |
关键词: Apoptosis; Proteases inhibitor; DNA digestion; Nuclear morphology; Etoposide; Mb; megabases; kb; kilobases; HMW; high molecular weight; FCS; foetal calf serum; PMC; puromycin; TBT; tributyltin; VP16; etoposide; TI; soy-bean trypsin inhibitor; PMSF; phenyl-methylsulphonyl-fluoride; IAA; iodoacetamide; NEM; N-ethyl-maleimide; TLCK; chloromethyl ketone; TPCK; chloromethyl ketone; | |
DOI : 10.1016/0014-5793(95)01284-2 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
The possible correlation between DNA digestion and changes in nuclear morphology in apoptosis was studied by blocking the apoptotic process at intermediate stages. The apoptogenic action of three drugs: etoposide, puromycin, tributyltin, was contrasted with protease inhibitors with different specificity on U937 cells. The inhibitors interfered with the development of the apoptotic features without shifting cell death to necrosis: treated cells showed abnormal morphologies, which could be recognized as intermediate stages of apoptosis; accordingly, DNA analysis showed an inhibitor-dependent block of the apoptotic DNA digestion. The comparison between size of DNA fragments and nuclear morphology suggested the following correlations: loss of normal nuclear shape with the appearence of a ≥ 2 Mb DNA band; ongoing chromatin condensation with the progressive DNA digestion up to 50 kb; nuclear fragmentation with DNA laddering. Protease inhibitors in etoposide-treated cells did not allow the formation of 700-300 kb fragments, suggesting that they possibly derive from a cell-mediated effect.
【 授权许可】
Unknown
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