Promoter methylation downregulates CDX2 expression in colorectal carcinomas

Hiroshi Kawai, Kunitoshi Tomii, Shinichi Toyooka, Masaaki Yano, Masakazu Murakami, Kazunori Tsukuda, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

CDX2 (caudal type homeobox transcription factor 2) is a homeobox protein, which is expressed in intestinal epithelium. CDX2 has been considered to play a role as a tumor suppressor gene in colorectal cancer because its expression is lacking in colorectal carcinomas, but preserved in adenomas. The point mutations have been found to account for loss of CDX2 expression, but the main mechanism responsible for CDX2 gene inactivation is less understood. We analyzed methylation and expression status of CDX2 in colorectal cancer cell lines and primary tumors. There are two CpG-rich sites in promoter region of CDX2 gene, -1570 to -1200 and -220 to +880. By COBRA (combined bisulfite restriction analysis) assay, the upper CpG-rich site was heavily methylated in all cell lines, but the lower CpG-rich site was methylated in limited cell lines. Bisulfite sequencing analysis and RT-PCR revealed that methylation of the lower CpG site was associated with down-regulation of CDX2. In addition, gene expression was restored in COLO201, a methylated cell line with 5-aza-2′-deoxycytidine, confirming that methylation caused gene down-regulation. We also examined CDX2 promoter methylation of primary tumors by MSP (methylated-allele specific PCR) assay and found that nearly 40% of cases have a methylated CDX2 gene. Our results demonstrate that CDX2 methylation is frequently present in colorectal cancers and may play a key role in inactivating CDX2 expression.

Original languageEnglish
Pages (from-to)547-551
Number of pages5
JournalOncology Reports
Volume13
Issue number3
Publication statusPublished - Mar 2005

Fingerprint

Methylation
Colorectal Neoplasms
Down-Regulation
decitabine
Cell Line
Genes
Homeodomain Proteins
Polymerase Chain Reaction
Gene Silencing
Intestinal Mucosa
Tumor Cell Line
Tumor Suppressor Genes
Point Mutation
Genetic Promoter Regions
Adenoma
Alleles
Gene Expression
Neoplasms
hydrogen sulfite

Keywords

  • Caudal type homeobox transcription factor 2
  • Colorectal carcinoma
  • DNA methylation

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Kawai, H., Tomii, K., Toyooka, S., Yano, M., Murakami, M., Tsukuda, K., & Shimizu, N. (2005). Promoter methylation downregulates CDX2 expression in colorectal carcinomas. Oncology Reports, 13(3), 547-551.

Promoter methylation downregulates CDX2 expression in colorectal carcinomas. / Kawai, Hiroshi; Tomii, Kunitoshi; Toyooka, Shinichi; Yano, Masaaki; Murakami, Masakazu; Tsukuda, Kazunori; Shimizu, Nobuyoshi.

In: Oncology Reports, Vol. 13, No. 3, 03.2005, p. 547-551.

Research output: Contribution to journalArticle

Kawai, H, Tomii, K, Toyooka, S, Yano, M, Murakami, M, Tsukuda, K & Shimizu, N 2005, 'Promoter methylation downregulates CDX2 expression in colorectal carcinomas', Oncology Reports, vol. 13, no. 3, pp. 547-551.
Kawai H, Tomii K, Toyooka S, Yano M, Murakami M, Tsukuda K et al. Promoter methylation downregulates CDX2 expression in colorectal carcinomas. Oncology Reports. 2005 Mar;13(3):547-551.
Kawai, Hiroshi ; Tomii, Kunitoshi ; Toyooka, Shinichi ; Yano, Masaaki ; Murakami, Masakazu ; Tsukuda, Kazunori ; Shimizu, Nobuyoshi. / Promoter methylation downregulates CDX2 expression in colorectal carcinomas. In: Oncology Reports. 2005 ; Vol. 13, No. 3. pp. 547-551.
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