BMC Cancer 2002, 2 http://www.biomedcentral.com/1471-2407/2/25 Table 1: Summary of clinicopathological data Tumor no. Sex/age Family history (No, Am, Fa, Ud) Tumor location Pre-op radio-therapy Dukes'/TNM stage Diff. grade Adjuvant Met.free/overall chemo-therapy survival (mo.) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 25/F 31/F 32/M 32/M 33/M 37/M 38/M 38/M 39/M 39/F 41/M 41/M 42/M 43/F 43/M 44/M 44/F 45/M 45/M 47/M 47/F 47/M 47/F 48/M 49/M 49/M 49/M 49/M 49/F 49/F No No Fa No No No No Am Ud Am No Fa Fa No No No Fa No No No Fa No No No No Ud No No No Fa m h m m m h m h l h h h l h m m m m h m l m l h h h h m m l + + + + + - + - + - + + + + + + - + + - + - + + + + - + + + C/T4N1M0 C/T3N1M0 D/T4N1M1 B/T3N0M0 B/T3N0M0 B/T3N0M0 A/T2N0M0 A/T1N0M0 D/T4N2M1 A/T2N0M0 A/T2N0M0 B/T3N0M0 C/T3N1M0 D/T4N2M1 C/T3N1M0 B/T3N0M0 C/T3N1M0 B/T3N0M0 C/T4N1M0 C/T3N1M0 C/T3N1M0 D/T3N0M1 C/T3N2M0 D/T4N2M1 D/T4N2M1 C/T3N1M0 C/T3N2M0 A/T2N0M0 A/T2N0M0 C/T3N1M0 mo mo mo mo mo mo mo we mu, si mo mu mo mo mo mo po mu mo mo mo mo mo mu, si mo mu mo mo mo mo mo FLV FLV FLV+OX FLV - - - - - - - FLV FLV FLV FLV - FLV - FLV - FLV FLV FLV FLV FLV FLV - - - - 46 84 0/15 33 85 80 61 49 0/8 32 35 34 27 0/20 46 19 10/22 71 55 28 24 0/40 11/17 0/24 0/3 27/38 62 37 72 71 Abbreviations: family history; Am: Amsterdam II criteria fulfilled, Fa: familial, No: no history; Ud:undetermined tumor location; h:high, m:middle, l:low differentiation; we:well, mo:moderate, po:poor, mu:mucinous, si:signet ring cell FLV; 5-FU + leucovorin performed on an Applied Biosystems 373 sequencer using cycle sequencing with AmpliTaq polymerase FS and the reagents supplied by Applied Biosystems (Foster City, CA, USA). MSI-analysis MSI was determined using the mononucleotide markers BAT25, BAT26, BAT34C4, BAT40 and the dinucleotide markers D2S123 and D17S250. All these MSI markers are among those recommended in the NCI reference panel for the classification of MSI [19]. All primer sequences are available from the authors upon request. Fluorescently la- belled forward primers were used for the PCR reactions: Tet (green) for BAT25, Fam (blue) for BAT26, BAT34C4 and D2S123 and Hex (yellow) for BAT40 and D17S250. The PCR products were mixed with formamide and Ge- neScan 500 TAMRA size standard (Applied Biosystems), denatured and separated in Performance Optimized Poly- mer-4 (POP-4™) on a ABI PRISM™ 310 Genetic Analyzer (Applied Biosystems). The tumors were classified as MSI- high if at least 2/5 markers displayed MSI, as MSI-low if instability was present in 1/5 markers and as MSS if none of the 5 markers showed instability. CTNNB1 mutations Exon 3 of the CTNNB1 gene, which codes for -catenin, was analysed using DNA sequencing and a PCR-based as- say designed to detect larger deletions of this region. Exon 3 was sequenced using the primers 5'-TGTAAAACGACG- GCCAGTCGTATTTATAGCTGATTTGA-3' (forward) and 5'-TACTCTTACCAGCTACTTGT-3' (reverse). Genomic de- letions involving exon 3 were analyzed by PCR amplific- tion using the primers 5'-CCAGCGTGGACAATGGCTAC- Page 3 of 8 (page number not for citation purposes)