Field Cancerization in Sporadic Colon Cancer

Abstract

Background/aims: Aberrant DNA methylation has a specific role in field cancerization. Certain molecular markers, including secreted frizzled-related protein 2 (SFRP2), tissue factor pathway inhibitor 2 (TFPI2 ), N-Myc downstream-regulated gene 4 (NDRG4) and bone morphogenic protein 3 (BMP3), have previously been shown to be hypermethylated in colorectal cancer (CRC). We aim to examine field cancerization in CRC based on the presence of aberrant DNA methylation in normal-appearing tissue from CRC patients.

Methods: We investigated promoter methylation in 34 CRC patients and five individuals with normal colonoscopy results. CRC patients were divided into three tissue groups: tumor tissue, adjacent and nonadjacent normal-appearing tissue. The methylation status (positive: methylation level >20%) of SFRP2 , TFPI2 , NDRG4 , and BMP3 promoters was investigated using methylation-specific PCR.

Results: The methylation frequencies of the SFRP2 , TFPI2 , NDRG4 and BMP3 promoters in tumor/adjacent/nonadjacent normal-appearing tissue were 79.4%/63.0%/70.4%, 82.4%/53.6%/60.7%, 76.5%/61.5%/69.2%, 41.2%/35.7%/50.0%, respectively. The methylation levels of the SFRP, TFPI2, NDRG4 and BMP3 promoters in tumor tissues were significantly higher than those in normal-appearing tissue (SFRP2, p=0.013; TFPI2, p<0.001; NDRG4, p=0.003; BMP3, p=0.001). No significant correlation was observed between the methylation levels of the promoters and the clinicopathological variables.

Conclusions: The field effect is present in CRC and affects both the adjacent and nonadjacent normal-appearing mucosa.

Keywords: Colorectal neoplasms; DNA methylation; Epigenomics; Field effect.

Fig. 1

Representative methylation-specific polymerase chain reaction of promoters in tissues. (a) SFRP2; (b) TFPI2; (c) NDRG4; and (d) BMP3. NC, negative control; PC, positive control; NN, nonadjacent normal-appearing tissue; AN, adjacent normal-appearing tissue; T, primary colorectal tumor tissue; SFRP2, secreted frizzled-related protein 2; TFPI2, tissue factor pathway inhibitor 2; NDRG4, N-Myc downstream-regulated gene 4; BMP3, bone morphogenic protein 3.

Fig. 2

Distribution of methylation level (%) for nonadjacent normal-appearing tissue (NN), adjacent normal-appearing tissue (AN), and primary colorectal tumor tissue (T) in (A) SFRP2, (B) TFPI2, (C) NDRG4, and (D) BMP3, respectively. SFRP2, secreted frizzled-related protein 2; TFPI2, tissue factor pathway inhibitor 2; NDRG4, N-Myc downstream-regulated gene 4; BMP3, bone morphogenic protein 3. *p<0.05.