Am. Yousef et al., Structural analysis of cDNA encoding thymidylate synthase in hepatic metastases of human colorectal tumors, INT J COL R, 16(5), 2001, pp. 318-325
The enzyme thymidylate synthase (TS) is an important target of 5-fluorourac
il (FUra) that is utilized for the treatment of disseminated colorectal can
cer. One determinant of clinical response to FUra-based therapy is TS expre
ssion, with high levels of expression being predictive of poor response. In
the present investigation the levels of immunoreactive TS were analyzed in
human colon metastases in the liver (n=11). The levels of TS ranged from 0
.30 to 4.60 pmol TS/g tissue. A good correlation was observed between the l
evels of immunoreactive TS and TS mRNA (n=6. r=0.69). Of the 11 metastases
analyzed, 5 exhibited relatively high levels of TS expression. Two metastas
es with high TS expression were obtained from patients who received adjuvan
t therapy with FUra. In 4 metastases with relatively high levels of TS expr
ession, TS gene copy number was analyzed. No evidence for amplification of
TS gene sequences was observed. The basis for the high levels of TS express
ion was examined by structural analysis of TS cDNA. No nucleotide sequence
differences were detected in the coding regions of the TS genes from the me
tastases. Mutations were detected at positions 961 and/or 1031 in the 3'-un
translated regions of the TS gene from the metastases; mutations at these s
ites were also detected in DNA isolated from normal colon mucosa (n=4) and
primary colorectal tumors (n=4). No correlation was observed between TS exp
ression and the nucleotide alterations at these positions. Polymorphism was
observed in the 5'-untranslated regions of the TS gene in hepatic metastas
es (n=6). A general trend was observed between the structure of the 5'-untr
anslated region of the gene and TS expression.