PMID- 1319346
OWN - NLM
STAT- MEDLINE
DCOM- 19920724
LR  - 20190707
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 201
IP  - 1
DP  - 1992 Jul
TI  - Changes in nuclear chromatin related to apoptosis or necrosis induced by the DNA 
      topoisomerase II inhibitor fostriecin in MOLT-4 and HL-60 cells are revealed by 
      altered DNA sensitivity to denaturation.
PG  - 184-91
AB  - The antitumor drug fostriecin (phosphotrienin, FST) has been reported to exert 
      its cytostatic and cytotoxic effects via inhibition of DNA topoisomerase II. The 
      sensitivity of human lymphocytic leukemic MOLT-4 and promyelocytic HL-60 leukemic 
      cells to a wide range of FST concentrations was studied by analyzing the cell 
      cycle-specific effects and changes in nuclear chromatin induced by this 
      inhibitor. The latter was evaluated by assaying the sensitivity of DNA in situ to 
      acid-induced denaturation cytofluorimetrically, with the use of the metachromatic 
      fluorochrome acridine orange (AO), which differentially stains double-stranded 
      and denatured DNA. The cytostatic effects were observed soon after addition of 
      FST (at concentrations of 1-30 microM for MOLT-4 cultures and 1-5 microM for 
      HL-60 cultures) as a perturbation of cell progression through S and G2 phases of 
      the cell cycle. Cell progression through the cycle was halted at greater than 30 
      microM FST in MOLT-4 cultures and at greater than 5 microM in HL-60 cultures; the 
      effect was instantaneous and affected all phases of the cycle, so that no changes 
      in the cell cycle distribution were apparent with increasing length of exposure 
      to the drug. Instead, at these high FST concentrations, immediate cytotoxic 
      effects became evident, manifesting either as cell apoptosis or necrosis. 
      Apoptosis was observed only in the case of HL-60 cells, at FST concentrations of 
      5-100 microM, and was characterized by markedly increased sensitivity of DNA to 
      denaturation combined with a decrease in overall DNA stainability, either with 
      the DNA-specific dye DAPI or with AO, indicative of the activation of endogenous 
      nucleases. Necrotic cell death was observed at FST concentrations of 1 mM and at 
      greater than 30 microM for HL-60 and MOLT-4 cells, respectively: in both cases 
      the overall DNA stainability, with either DAPI or AO, was unchanged compared to 
      the control, but their DNA was very sensitive to denaturation. Interestingly, DNA 
      in G2 and late S phase MOLT-4 cells, which were undergoing necrotic death, was 
      much more sensitive to denaturation than was DNA in G1 cells of this lineage. The 
      data indicate that chromatin changes induced by DNA topoisomerase II inhibitors 
      in cells that undergo apoptotic or necrotic death can be conveniently monitored 
      by the assay of DNA in situ sensitivity to denaturation.
FAU - Hotz, M A
AU  - Hotz MA
AD  - Cancer Research Institute, New York Medical College, Valhalla 10595.
FAU - Traganos, F
AU  - Traganos F
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
LA  - eng
GR  - CA 28704/CA/NCI NIH HHS/United States
GR  - R37 CA 23296/CA/NCI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - 0 (Alkenes)
RN  - 0 (Chromatin)
RN  - 0 (Polyenes)
RN  - 0 (Pyrones)
RN  - 0 (Topoisomerase II Inhibitors)
RN  - F30N4O6XVV (Acridine Orange)
RN  - ZO1648L551 (fostriecin)
SB  - IM
MH  - Acridine Orange
MH  - Alkenes/pharmacology
MH  - Cell Death/drug effects
MH  - Cell Line/drug effects/enzymology/pathology
MH  - Cell Nucleus/chemistry/drug effects
MH  - Chromatin/*chemistry
MH  - G2 Phase/drug effects
MH  - Humans
MH  - Leukemia/drug therapy/pathology
MH  - Polyenes
MH  - Pyrones
MH  - S Phase/drug effects
MH  - *Topoisomerase II Inhibitors
MH  - Tumor Cells, Cultured/drug effects
EDAT- 1992/07/01 00:00
MHDA- 1992/07/01 00:01
CRDT- 1992/07/01 00:00
PHST- 1992/07/01 00:00 [pubmed]
PHST- 1992/07/01 00:01 [medline]
PHST- 1992/07/01 00:00 [entrez]
AID - 0014-4827(92)90362-C [pii]
AID - 10.1016/0014-4827(92)90362-c [doi]
PST - ppublish
SO  - Exp Cell Res. 1992 Jul;201(1):184-91. doi: 10.1016/0014-4827(92)90362-c.