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Titolo:
THE EFFECTS OF 5-FLUOROPYRIMIDINES ON NASCENT DNA-SYNTHESIS IN CHINESE-HAMSTER OVARY CELLS MONITORED BY PH-STEP ALKALINE AND NEUTRAL ELUTION
Autore:
JONES S; WILLMORE E; DURKACZ BW;
Indirizzi:
UNIV NEWCASTLE UPON TYNE,SCH MED,CANC RES UNIT NEWCASTLE TYNE NE2 4HHTYNE & WEAR ENGLAND
Titolo Testata:
Carcinogenesis
fascicolo: 11, volume: 15, anno: 1994,
pagine: 2435 - 2438
SICI:
0143-3334(1994)15:11<2435:TEO5ON>2.0.ZU;2-U
Fonte:
ISI
Lingua:
ENG
Soggetto:
MOUSE FM3A CELLS; THYMINELESS DEATH; HEPATOMA-CELLS; 5-FLUOROURACIL; 5-FLUORODEOXYURIDINE; TRIPHOSPHATE; CYTOTOXICITY; PURIFICATION; MECHANISMS; RESISTANCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
23
Recensione:
Indirizzi per estratti:
Citazione:
S. Jones et al., "THE EFFECTS OF 5-FLUOROPYRIMIDINES ON NASCENT DNA-SYNTHESIS IN CHINESE-HAMSTER OVARY CELLS MONITORED BY PH-STEP ALKALINE AND NEUTRAL ELUTION", Carcinogenesis, 15(11), 1994, pp. 2435-2438

Abstract

Nascent DNA (nDNA) replication intermediates can be isolated and quantified by pH-step alkaline elution (L.C. Erikson et al,, Chromosoma, 74, 125-139, 1979). The effects of 5-fluorouracil (FU) and 5-fluoro-2'-deoxyuridine (FdU) on the formation and persistence of nascent DNA structures were studied in Chinese hamster ovary K1 cells. Exogenous thymidine (dT) rescued FdU-induced, but had little effect on FU-induced, cytotoxicity, indicating DNA- and RNA-directed cytotoxic mechanisms respectively. Drug-treated cells were pulse labelled with [H-3]dT and thepercentage of total counts eluting from and retained by filters estimated at successive pH increments (11.0, 11.3, 11.5 and 12.1). A 1 min pulse of control cells resulted in >75% of the DNA eluting at pH 12.1 or being retained by the filter. By 1 h, >90% of the DNA was retained by the filter. FdU treatment resulted in the persistence of DNA in allfour pH bands. More than 95% of total DNA synthesized during a 5 min pulse eluted as discrete peaks at each pH, This DNA was processed to higher M(r) species during a 1 h chase, but following a 24 h chase, 68%of the DNA still eluted at pH 12.1. In contrast, FU treatment caused only a transient accumulation (less than or equal to 10 min) of the DNA (e.g. similar to 26% at 5 min) in the pH 11.0 and 11.3 bands only, indicating a selective effect of FU on the maturation of very short size classes of DNA. Neutral elution was used to assess the effect of FdUon double-strand break (dsb) formation in nDNA. Whereas no dsbs were evident in untreated cells, dsbs appeared in FdU-treated cells even following the shortest [H-3]dT pulse times (1-10 min). Although these were processed into higher M(r) species with longer pulse times, dsbs were still evident at 2 h.

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Documento generato il 01/10/20 alle ore 15:18:42