Trifluridine (FTD) is a key component of the novel oral
antitumor drug TAS-102, which consists of FTD and a
thymidine phosphorylase inhibitor. Like
5-fluoro-2'-deoxyuridine (FdUrd), a deoxynucleoside form of
5-fluorouracil metabolite, FTD is sequentially phosphorylated and not only inhibits
thymidylate synthase activity, but is also incorporated into
DNA. Although
TAS-102 was effective for the treatment of refractory metastatic
colorectal cancer in clinical trials, the mechanism of FTD-induced cytotoxicity is not completely understood. Here, we show that FTD as well as FdUrd induce transient phosphorylation of Chk1 at Ser345, and that this is followed by accumulation of p53 and p21
proteins in p53-proficient human
cancer cell lines. In particular, FTD induced p53-dependent sustained arrest at G2 phase, which was associated with a
proteasome-dependent decrease in the
Cyclin B1 protein level and the suppression of CCNB1 and CDK1 gene expression. In addition, a p53-dependent increase in p21
protein was associated with an FTD-induced decrease in
Cyclin B1 protein. Although numerous ssDNA and dsDNA breaks were induced by FdUrd, few
DNA strand breaks were detected in FTD-treated HCT-116 cells despite massive FTD misincorporation into genomic
DNA, suggesting that the antiproliferative effect of FTD is not due to the induction of
DNA strand breaks. These distinctive effects of FTD provide insights into the cellular mechanism underlying its antitumor effect and may explain the clinical efficacy of
TAS-102.