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concomitant absence of transcript <strong>and</strong> prote<strong>in</strong>occurs <strong>in</strong> small patches of crypt cells <strong>in</strong> unaffectedmucosa from sporadic colorectal carc<strong>in</strong>oma. DiagnMol Pathol 2006, 15:17-23.64. Ide T, Kitajima Y, Ohtaka K, Mitsu<strong>no</strong> M, Nakafusa Y,Miyazaki K: Expression of the hMLH1 gene is apossible predictor for the cl<strong>in</strong>ical response to 5-fluorouracil after a surgical resection <strong>in</strong> colorectalcancer. Oncol Rep 2008, 19:1571-1576.65. Hitch<strong>in</strong>s MP, L<strong>in</strong> VA, Buckle A, Cheong K, Halani N, KuS, Kwok CT, Packham D, Suter CM, Meagher A et al.:Epi<strong>genetic</strong> <strong>in</strong>activation of a cluster of genesflank<strong>in</strong>g MLH1 <strong>in</strong> microsatellite-unstable colorectalcancer. Cancer Res 2007, 67:9107-9116.66. Leung WK, To KF, Man EP, Chan MW, Bai AH, Hui AJ,Chan FK, Sung JJ: Quantitative detection ofpromoter hypermethylation <strong>in</strong> multiple genes <strong>in</strong> theserum of patients with colorectal cancer. Am JGastroenterol 2005, 100:2274-2279.67. Xiong Z, Wu AH, Bender CM, Tsao JL, Blake C,Shibata D, Jones PA, Yu MC, Ross RK, Laird PW:Mismatch repair deficiency <strong>and</strong> CpG isl<strong>and</strong>hypermethylation <strong>in</strong> sporadic colonade<strong>no</strong>carc<strong>in</strong>omas. Cancer Epidemiol Biomarkers Prev2001, 10:799-803.68. Muller HM, Oberwalder M, Fiegl H, Mor<strong>and</strong>ell M,Goebel G, Zitt M, Muhlthaler M, Ofner D, Margreiter R,Widschwendter M: Methylation changes <strong>in</strong> faecalDNA: a marker for colorectal cancer screen<strong>in</strong>g?Lancet 2004, 363:1283-1285.69. Chen WD, Han ZJ, Skoletsky J, Olson J, Sah J, MyeroffL, Platzer P, Lu S, Dawson D, Willis J et al.: Detection<strong>in</strong> fecal DNA of colon cancer-specific methylatio<strong>no</strong>f the <strong>no</strong>nexpressed viment<strong>in</strong> gene. J Natl CancerInst 2005, 97:1124-1132.70. Giovannucci E, Og<strong>in</strong>o S: DNA methylation, fieldeffects, <strong>and</strong> colorectal cancer. J Natl Cancer Inst2005, 97:1317-1319.71. Fe<strong>in</strong>berg AP, Ohlsson R, Henikoff S: The epi<strong>genetic</strong>progenitor orig<strong>in</strong> of human cancer. Nat Rev Genet2006, 7:21-33.Additional materialAdditional file 1:File format: DOCTitle: Supplementary <strong>in</strong>formationDescription: Methodological details which are <strong>no</strong>t crucial forthe underst<strong>and</strong><strong>in</strong>g of the work, as well as Additional figurelegends.Additional file 2:File format: JPGTitle: Titration of methylated DNA template illustrates thescor<strong>in</strong>g thresholds for the methylation-specific polymerasecha<strong>in</strong> reaction.Description: Determ<strong>in</strong>ation of scor<strong>in</strong>g thresholds is visualizedby a titration series of the RUNX3 gene.Additional file 3:File format: XLSTitle: PCR primers used for methylation-specific PCR <strong>and</strong>microsatellite <strong>in</strong>stability analyses.Description: Information on primers <strong>and</strong> PCR detailsAdditional file 4:File format: XLSTitle: Genetic <strong>and</strong> epi<strong>genetic</strong> raw data.Description: Raw data from all analyses are listed for eachtumor <strong>in</strong>cluded.Additional file 5:File format: XLSTitle: Summarized methylation raw data.Description: Methylation frequencies are presented for eachof the eleven analyzed genes <strong>in</strong> the 4 different sample typesaccord<strong>in</strong>g to what methylation level they exhibited (strong,weak or <strong>no</strong> methylation).Additional file 6:File format: JPGTitle: Comparison between MSP <strong>and</strong> quantitative MSP <strong>in</strong><strong>no</strong>rmal mucosa samples.Description: Hypermethylation of was analyzed with both<strong>no</strong>n-quantitative- <strong>and</strong> quantitative MSP. Here the results fromeach method are presented.Additional file 7:File format: XLSTitle: Correlation between methylated genes.Description: A correlation table <strong>in</strong>clud<strong>in</strong>g all analyzed forpromoter hypermethylation shown that genes commonlymethylated <strong>in</strong> MSI tumors are highly correlated.Additional file 8:File format: JPGTitle: Widespread methylation among <strong>no</strong>rmal colorectalsamples, ade<strong>no</strong>mas, <strong>and</strong> carc<strong>in</strong>omas.Description: A histogram show<strong>in</strong>g the total number ofmethylated genes per sample <strong>in</strong> <strong>no</strong>n-cancerous <strong>no</strong>rmalmucosa, <strong>no</strong>rmal mucosa taken <strong>in</strong> distance from a primarytumor, ade<strong>no</strong>mas, <strong>and</strong> carc<strong>in</strong>omas stratified accord<strong>in</strong>g toMSI status.10

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