Mitotic Arrest Induced By Xk469, A Novel Antitumor Agent, Is Correlated With The Inhibition Of Cyclin B1 Ubiquitination

XK469 (NSC 697887) is a novel antitumor agent with broad activity against a variety of tumors. Previous studies suggest that XK469 is a topoisomerase lip poison with functional activity similar to that of 4'- (9-acridinylamino) methanesulfon-m-anisidide (m-AMSA). The goal of our study was to investigate its mechanism of action further using a human HCT-116 (H116) colon tumor cell model. Concentration-survival curves with continuous exposure indicated that XK469 had low cytotoxic activity against H116 cells. Cell cycle analysis revealed that XK469 is a phase-specific cell cycle blocker that is associated with increased levels of cyclin BI, cyclin A and p53 but not CDKI (cdc2) or cyclin E. In contrast, treatment of H116 cells with m-AMSA caused a total degradation of both cyclin A and BI but enhanced expression of cyclin E and p53. Accumulation of cyclin BI in XK469-treated cells was correlated with the inhibition of cyclin BI ubiquitination, a metabolic process mandatory for proteasome-mediated protein turnover. However, no inhibition of cyclin BI ubiquitination was detected in cells treated with rn-AMSA or colchicine, a known mitotic inhibitor. Furthermore, unlike rn-AMSA, XK469 did not induce caspase activation or apoptotic cell death in H116 cells. Our results suggest that XK469 is a phase-specific cell cycle inhibitor with a unique mechanism of action that is correlated with the inhibition of cyclin BI ubiquitination and its accumulation at early M phase. (C) 2002 Wiley-Liss, Inc.