Thymidylate synthetase (TS) genotype and TS/dihydropyrimidine dehydrogenase mRNA level as an indicator in determining chemosensitivity to 5-fluorouracil in advanced gastric carcinoma.
BACKGROUND
One of the target enzymes of 5-fluorouracil (5-FU) is thymidylate synthetase (TS). The DNA sequence of the TS gene includes either double or triple tandem 28-bp repeats within the promoter region, such that TS genotypes can be classified as homozygous 3R/3R heterozygous 2R/3R or homozygous 2R/2R Several recent studies have shown that TS genotype affects mRNA expression, with 3R/3R homozygotes showing higher TS mRNA expression compared to the other genotypes.


OBJECTIVE
We analyzed the TS genotype and TS and dihydropyrimidine dehydrogenase (DPD) mRNA expression levels in 22 advanced gastric carcinoma patients, and analyzed results with respect to patient 5-FU chemosensitivity, as detected by the tetrazolium-based colorimetric (MTT) assay and survival outcome.


METHODS
Between September 2001 and April 2002, 22 Japanese patients with advanced gastric carcinoma were evaluated. Informed written consent was obtained from all patients and the study was approved by the ethical committee at our University Hospital Fresh surgical specimens from carcinoma lesions were enzymatically dissociated and incubated with 5-FU at a concentration of 50 microg/ml for 48 hours to determine the inhibition rate as detected by MTT assay. Normal and tumor tissue and peripheral blood samples were collected and stored at -80 degrees C until assay for TS genotype and TS and DPD mRNA level The TS genotype was assessed by PCR assay using peripheral monocytes, since monocyte genotypes represent the genotype of normal and tumor tissues. Quantification of TS and DPD mRNA levels was performed using real-time PCRP Survival outcome was assessed according to the disease-free survival period in cases with similar clinical backgrounds.


RESULTS
TS genotyping revealed 19 3R/3R homozygotes and 3 2R/3R heterozygotes. After analysis of normal and tumor tissues, samples from homozygote 3R/3R cases showed higher TS mRNA expression than heterozygote 2R/3R cases, which was statistically significant at p<0.05. We also observed a statistically significant correlation in TS mRNA levels between normal and tumor tissues, while no significant correlation was observed for DPD mRNA levels between normal and tumor tissues. While no relationship between 5-FU chemosensitivity and TS genotype or mRNA expression was observed, cases with high DPD mRNA expression were resistant to 5-FU and exhibited poor survival outcomes.


CONCLUSIONS
While TS genotype affected TS mRNA expression in both normal and tumor tissues in advanced gastric cancer, there is no relationship between TS genotype or mRNA expression level and 5-FU chemosensitivity.


CONCLUSIONS
Our finding, that DPD mRNA expression appears to be a factor in determining 5-FU chemosensitivity and the survival outcome of advanced gastric cancer patients, is comparable with previous reports.
Department of Surgery, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Fumiki
F
Toriumi
Tetsuro
T
Kubota
Yoshiro
Y
Saikawa
Masashi
M
Yoshida
Yoshihide
Y
Otani
Masahiko
M
Watanabe
Koichiro
K
Kumai
Masaki
M
Kitajima
D000368:Aged; D000369:Aged, 80 and over; D000964:Antimetabolites, Antineoplastic / Q000493:pharmacokinetics / Q000494:pharmacology*; D001483:Base Sequence; D003124:Colorimetry / Q000379:methods; D042943:Dihydrouracil Dehydrogenase (NADP) / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004338:Drug Combinations; D005260:Female; D005472:Fluorouracil / Q000493:pharmacokinetics / Q000494:pharmacology*; D005838:Genotype; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000201:enzymology / Q000502:physiology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010094:Oxonic Acid / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000188:drug therapy / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D005641:Tegafur / Q000494:pharmacology; D013778:Tetrazolium Salts; D013844:Thiazoles; D013940:Thymidylate Synthase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
D000964:Antimetabolites, Antineoplastic; D004338:Drug Combinations; D011725:Pyridines; D012333:RNA, Messenger; D013778:Tetrazolium Salts; D013844:Thiazoles; C079198:S 1 (combination); D005641:Tegafur; D010094:Oxonic Acid; D042943:Dihydrouracil Dehydrogenase (NADP); D013940:Thymidylate Synthase; C022616:thiazolyl blue; D005472:Fluorouracil