No cases of NN displayed TaqMeth V over the optimal

No cases of NN displayed TaqMeth V over the optimal cut-offs for B4GALT1 and OSMR (100% specificity). The sensitivities of B4GALT1 and OSMR were 83% (25/30) and 80% (20/25), respectively. These results indicate that B4GALT1 and OSMR harbor cancer-specific methylation in CRC with high frequency. Therefore, we focused on B4GALT1 and OSMR for further study. We examined the methylation status of B4GALT1 and OSMR in 100 new pairs of CRC (PT) and corresponding normal (PN) tissues by TaqMan-MSP analysis (Fig. 1C). The TaqMeth V in PT ranged from 0 to 370.70 (median value 3.70) for B4GALT1 and from 0 to 749.27 (median value 59.24) for OSMR. The value in PN ranged from 0 to 9.70 (median value 0.26) for B4GALT1 and to 190.68 (median value 1.54) for OSMR. The overall methylation levels of B4GALT1 and OSMR detected in PT (22.

33��48.69 for B4GALT1 and 116.83��151.52 for OSMR, mean��SD, n=100) were also significantly higher than those in PN (0.90��1.37 for B4GALT1 and 6.49��21.52 for OSMR, mean��SD, n=100) (P<0.001). At the optimal cut-offs (values, 3.87 for B4GALT1 and 22.01 for OSMR) calculated from the ROC analysis (PT vs. PN) (Figure S4B), the specificity of the two genes was over 96%. The sensitivities of B4GALT1 and OSMR were 49% (49/100) and 80% (80/100), respectively. Taken together, B4GALT1 and OSMR were frequently methylated in primary CRC tissues but displayed absent or low levels of methylation in corresponding normal tissues. Next, we performed a blinded analysis of gene methylation analysis in stool DNA collected from patients with or without colon cancer.

The clinical status of the patients was not revealed until the analysis was completed. We also examined hypermethylation of SFRP1 as a positive control for detection of gene methylation in stool DNA [18]. The results of ROC analysis in the stool DNA are shown in Figure 2. Table 4 shows the sensitivity/specificity of each gene for colon cancer detected in stool DNA samples. SFRP1 methylation was not detected in patients with endoscopically normal colon (0%, 0/15), and was found in 55% (11/20) of CRC patients, at the optimal cut-off value calculated from the ROC analysis. Methylation of B4GALT1 and OSMR was detected in 64% (9/14) and 80% (16/20) of stool DNA samples from CRC patients, respectively. When methylation of SFRP1 and OSMR was combined, the sensitivity was 60% (12/20), and the specificity was 100% (0/15).

Discrimination between patients without cancer and CRC patients by SFRP1 or OSMR methylation was statistically significant (P<0.01). Figure 2 ROC curve Carfilzomib analysis in stool DNA. Table 4 Gene methylation detected in stool from colon cancer or non-cancer patients. OSMR methylation was tested in a blinded fashion in one more independent set of stool samples (no overlap with the samples in Table 4). Stool DNA from healthy control subjects who had no visual abnormalties in colonoscopy were included for this study.

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