PMC

The percentage of ADHFE1 methylated reference of CRC lines. All CRC cell lines showed more than 10% PMR. The ADHFE1 promoter was hypermethylated in all CRC cell lines.

The bisulfite genomic sequencing of the ADHFE1 promoter region. Filled (black) circles correspond to the methylated CpG Island, unfilled (white) circles correspond to unmethylated CpG Island.

Localization of ADHFE1 in normal colorectal mucosa (×200). (A) The ADHFE1 expression was detected predominantly in the apex of the crypts. It was noted to be decreased toward the bottom of the crypts. (B) A strong expression of ADHFE1 can be observed in the Paneth cells (arrow).

The overexpression of ADHFE1 induces the expression of a colon differentiation marker. (A) The ALP activity was increased in the DLD-1 cell line transfected with pcDNA3.1 vector containing the coding region of ADHFE1, compared to controls. (B) The expression of ADHFE1, CEA and Cdx2 is increased in the DLD-1 cell line transfected with pcDNA3.1 vector containing the coding region of ADHFE1 and controls.

The expression of ADHFE1 protein in CRC. A western blotting analysis was performed to examine ADHFE1 protein expression in CRC (T) and adjacent normal colorectal tissue (N). These 8 blots are representative. The corresponding reprobings of these blots with actin as a loading control.

ADHFE1 promoter methylation and mRNA expression with and without treatment with 5-aza-2-dC. (A) After treatment of 5-aza-2-dC, the relative methylation of ADHFE1 promoter was decreased in 9 of 12 CRC cell lines. (B) After treatment of 5-aza-2-dC, the expression of ADHFE1 mRNA was increased in 11 of 12 CRC cell lines.

The expression of ADHFE1 during mouse intestinal differentiation and development. (A) A broad and diffuse staining of ADHFE1 is found in the early stage of gut at E15.5 (×5). (B) With progressive villous formation, ADHFE1 is expressed along the whole anterior-posterior axis of epithelium at E17.5 (×5). (C) At E18.5, ADHFE1 expression is more delineated in the top of the epithelium as differentiation proceeds (×20).

The expression of ADHFE1 according to the differentiation status in CRC. (A-C) Good differentiation is associated with the strong expression of ADHFE1. (A), well differentiated CRC; (B), moderate differentiated CRC; (C), poorly differentiated CRC (×5). (D) There is a correlation between ADHFE1 expression and differentiation status in CRC. (E) A representative case shows a simultaneously taken picture with a different gradient of differentiation in CRC (×100). MD, moderate differentiated CRC; WD, well differentiated CRC.