The published data about thymidylate synthase (TS) expression and its predictive value in advanced colorectal cancer (CRC) patients receiving fluoropyrimidine-based chemotherapy seemed inconclusive. To derive a more precise estimation of the relationship, a metaanalysis was performed. Studies have been identified by searching PubMed and Embase. Inclusion criteria were advanced CRC patients, received fluoropyrimidine-based chemotherapy and evaluation of TS expression and overall response rate (ORR). The relative ratio (RR) for ORR in patients with low-TS expression over those with high-TS expression with 95% confidence interval (CI) was calculated for each study as an estimation of the predictive effect of TS. A total of 24 studies including 1,112 patients were involved in this metaanalysis. The overall RR was 2.20 (95% CI, 1.82-2.66; p 5 0.000). For studies evaluating TS expression in metastatic lesions, the pooled RR was 3.23 (95% CI, 2.27-4.59; p 5 0.000); for studies testing TS expression in primary lesions, a pooled RR of 1.89 (95% CI, 1.45-2.48; p 5 0.000) was estimated. Focusing the analysis on immunohistochemistry (IHC)-based or RTPCR-based assessments, the pooled RR was 1.83 (95% CI, 1.44-2.34; p 5 0.000) and 2.96 (95% CI, 2.07-4.22; p 5 0.000), respectively. The results indicated that low-TS expression tumors in advanced CRC patients were more sensitive to fluoropyrimidine-based chemotherapy. Subgroup analyses indicated that the predictive value of TS expression evaluated in metastases was more prominent than that of primary lesions, and that TS expression tested by RTPCR was also of greater predictive value than by IHC. ' 2008 Wiley-Liss, Inc.Key words: thymidylate synthase; predictive value; advanced colorectal cancer; fluoropyrimidine-based chemotherapy; metaanalysis Colorectal cancer (CRC) is the 3rd most common malignant disease and the 4th most frequent cause of cancer-related deaths worldwide, with an estimated 1 million new cases and 0.5 million deaths every year. In developed countries, CRC is the 2nd most common tumor with a lifetime incidence of 5%. The prognosis of CRC is poor, with about half of all diagnosed patients dying of metastatic spread. In the advanced setting, the mainstay of treatment remains fluoropyrimidine-based chemotherapy. 1,2 Unfortunately, some patients do not benefit from fluoropyrimidine-based treatment strategies. Therefore, predictive factors are needed to identify the subgroup that is most likely to profit from such chemotherapy protocols. Thymidylate synthase (TS), which is an important enzyme for DNA synthesis, is the target of fluoropyrimidine. 3 The main mechanism of fluoropyrimidines antitumor effect is ascribed to be the competitive inhibition of TS after conversion to its active metabolite. 4 TS expression as a determinant of sensitivity to fluoropyrimidines has been demonstrated in vitro, 5,6 whereas TS expression in vivo has attracted a considerable attention because of its potential role as a promising predictive factor for response to fluoropyrimidine-base...