J. Ma et al., REDUCED CELLULAR ACCUMULATION OF TOPOTECAN - A NOVEL MECHANISM OF RESISTANCE IN A HUMAN OVARIAN-CANCER CELL-LINE, British Journal of Cancer, 77(10), 1998, pp. 1645-1652
In order to unravel possible mechanisms of clinical resistance to topo
isomerase I inhibitors, we developed a topotecan-resistant human IGROV
-1 ovarian cancer cell line, denoted IGROV(T100r), by stepwise increas
ed exposure to topotecan (TPT). The IGROV(T100r) cell line was 29-fold
resistant to TPT and strongly cross-resistant to SN-38 (51-fold). How
ever, the IGROV(T100r) showed only threefold resistance to camptotheci
n (CPT). Remarkably, this cell line was 32-fold resistant to mitoxantr
one, whereas no significant cross-resistance against other cytostatic
drugs was observed. No differences in topoisomerase I protein levels a
nd catalytic activity as well as topoisomerase I cleavable complex sta
bilization by CPT in the IGROV-1 and IGROV(T100r) cell lines were obse
rved, indicating that resistance in the IGROV(T100r) cell line was not
related to topoisomerase I-related changes. However, resistance in th
e resistant IGROV(T100r) cell line to TPT and SN-38 was accompanied by
decreased accumulation of the drugs to approximately 15% and 36% of t
hat obtained in IGROV-1 respectively No reduced accumulation was obser
ved for CPT. Notably, accumulation of TPT in the IGROV-1 cell line dec
reased under energy-deprived conditions, whereas the accumulation in t
he IGROV(T100r) cell line increased under these energy-deprived condit
ions. The efflux of TPT at 37 degrees C was very rapid in the IGROV-1
as well as the IGROV(T100r) cell line, resulting in 90% efflux within
20 min. Importantly, the efflux rates of TPT in the IGROV-1 and IGROV(
T100r) cell lines were not significantly different and were shown to b
e independent on P-glycoprotein (P-gp) or multidrug resistance-associa
ted protein (MRP). These results strongly suggest that the resistance
of the IGROV(T100r) cell line to TPT and SN-38 is mainly caused by red
uced accumulation. The reduced accumulation appears to be mediated by
a novel mechanism, probably related to impaired energy-dependent uptak
e of these topoisomerase I drugs.