J Korean Med Sci.  2010 Mar;25(3):405-417. 10.3346/jkms.2010.25.3.405.

DNA Methylation Patterns of Ulcer-Healing Genes Associated with the Normal Gastric Mucosa of Gastric Cancers

• Hong SJ
• Oh JH
• Jung YC
• Kim YH
• Kim SJ
• Kang SJ
• Seo EJ
• Choi SW
• Kang MI
• Rhyu MG

• KMID: 1778036
• DOI: http://doi.org/10.3346/jkms.2010.25.3.405

Abstract

Recent evidence suggests that gastric mucosal injury induces adaptive changes in DNA methylation. In this study, the methylation status of the key tissue-specific genes in normal gastric mucosa of healthy individuals and cancer patients was evaluated. The methylation-variable sites of 14 genes, including ulcer-healing genes (TFF1, TFF2, CDH1, and PPARG), were chosen from the CpG-island margins or non-island CpGs near the transcription start sites. The healthy individuals as well as the normal gastric mucosa of 23 ulcer, 21 non-invasive cancer, and 53 cancer patients were examined by semiquantitative methylation-specific polymerase chain reaction (PCR) analysis. The ulcer-healing genes were concurrently methylated with other genes depending on the presence or absence of CpG-islands in the normal mucosa of healthy individuals. Both the TFF2 and PPARG genes were frequently undermethylated in ulcer patients. The over- or intermediate-methylated TFF2 and undermethylated PPARG genes was more common in stage-1 cancer patients (71%) than in healthy individuals (10%; odds ratio [OR], 21.9) and non-invasive cancer patients (21%; OR, 8.9). The TFF2-PPARG methylation pattern of cancer patients was stronger in the older-age group (> or =55 yr; OR, 43.6). These results suggest that the combined methylation pattern of ulcer-healing genes serves as a sensitive marker for predicting cancer-prone gastric mucosa.

Keyword

DNA Methylation; Stomach; Ulcer; Non-Invasive Cancer; Neoplasms

MeSH Terms

Biological Markers/metabolism
Cadherins/genetics
CpG Islands
*DNA Methylation
Female
*Gastric Mucosa/pathology/physiology
Gene Expression Regulation, Neoplastic
Growth Substances/genetics
Humans
Male
Middle Aged
Neoplasm Invasiveness
PPAR gamma/genetics
Peptides/genetics
*Stomach Neoplasms/genetics/pathology
*Stomach Ulcer/genetics/pathology
Tumor Suppressor Proteins/genetics
Wound Healing/*genetics
Biological Markers
Cadherins
Growth Substances
PPAR gamma
Peptides
Tumor Suppressor Proteins

Figure

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  Fig. 1 Schematic diagram of the length of the CpG-islands and the retroelement distributions in the 5'-end regions of 14 selected genes (A) and their serial analysis of gene expression (SAGE) profiles in the normal gastric mucosa and gastric cancer tissue (B).

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  Fig. 2 Representative autoradiographs and ethidium bromide staining of electrophoresis bands generated from both unmethylation (U) and methylation (M) PCR. Genomic DNA was obtained from normal mucosa of normal control (A), gastric ulcer (B), non-invasive cancer (C), and stage-1 cancer (D) patients.

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  Fig. 3 The methylation status of the methylation-variable sites in the normal mucosa of normal stomachs (A) and 10 bone marrow stromal cells (B).

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  Fig. 4 Methylation patterns of ulcer-healing (TFF1, TFF2, and CDH1) genes in the normal gastric mucosa analyzed according to the age groups (A) and H. pylori infection (B).

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  Fig. 5 Methylation-variations in the normal mucosa of stage-1 and advanced-stage gastric cancer patients (A) and comparison of the 6 selected genes examined in normal mucosa samples collected 2 cm and 5 cm distal from pathologic lesions (B). Asterisk indicates statistically significant.

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  Fig. 6 Concurrent methylation status between the ulcer-healing (CDH1, TFF1, and TFF2) genes and the other genes in the normal gastric mucosa of healthy individuals and stage-1 gastric cancer patients. Correlation coefficients (R) were calculated by Pearson's correlation. Asterisk indicates statistically significant. Asterisk indicates statistically significant.

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