Dj. Grdina et al., INHIBITION OF TOPOISOMERASE II-ALPHA ACTIVITY IN CHO K1 CELLS BY 2-[(AMINOPROPYL)AMINO]ETHANETHIOL (WR-1065), Radiation research, 138(1), 1994, pp. 44-52
The aminothiol 2-[(aminopropyl)amino]ethanethiol (WR-1065) is the acti
ve thiol of the clinically studied radioprotective agent S-2-(3-aminop
ropylamino)ethylphosphorothioic acid (WR-2721). WR-1065 is an effectiv
e radiation protector when it is administered 30 min prior to exposure
of Chinese hamster ovary K1 cells to radiation (i.e., a dose modifica
tion factor of 1.4) at a concentration of 4 mM. Under these exposure c
onditions, topoisomerase (Topo) I and II alpha activities and associat
ed protein contents-were measured in cells of the K1 cell line using t
he DNA relaxation assay, the P4 unknotting assay and immunoblotting, r
espectively. WR-1065 was ineffective in modifying Topo I activity, but
it did reduce Topo II alpha activity by an average of 50%. The magnit
ude of Topo II alpha protein content, however, was not affected by the
se exposure conditions. The effects on the cell cycle were monitored b
y the method of flow cytometry. Exposure of cells to 4 mM WR-1065 for
up to 6 h resulted in a build-up of cells in the G(2)/M-phase compartm
ent. However, under these conditions and in contrast to Topo II inhibi
tors used in chemotherapy, WR-1065 is an effective radioprotective age
nt capable of protecting against both radiation-induced cell lethality
and mutagenesis. One of several mechanisms of action attributed to am
inothiol compounds such as WR-1065 has been their ability to affect en
dogenous enzymatic reactions involved in DNA synthesis and repair and
progression of cells through the phases of the cell cycle. These resul
ts are consistent with such a proposed mechanism and demonstrate in pa
rticular a modifying effect by WR-1065 on Topo II, which is involved i
n DNA synthesis.