Cancer Res Treat.  2019 Jan;51(1):267-279. 10.4143/crt.2018.085.

Gastric Mucosal Atrophy Impedes Housekeeping Gene Methylation in Gastric Cancer Patients

• Oh JH
• Rhyu MG
• Kim SI
• Yun MR
• Shin JH
• Hong SJ

• KMID: 2437618
• DOI: http://doi.org/10.4143/crt.2018.085

Abstract

PURPOSE
Helicobacter pylori infection induces phenotype-stabilizing methylation and promotes gastric mucosal atrophy that can inhibit CpG-island methylation. Relationship between the progression of gastric mucosal atrophy and the initiation of CpG-island methylation was analyzed to delineate epigenetic period for neoplastic transformation.
MATERIALS AND METHODS
Normal-appearing gastric mucosa was biopsied from 110 H. pylori-positive controls, 95 H. pylori-negative controls, 99 gastric cancer patients, and 118 gastric dysplasia patients. Gastric atrophy was assessed using endoscopic-atrophic-border score. Methylation-variable sites of eight CpG-island genes adjacent to Alu (CDH1, ARRDC4, PPARG, and TRAPPC2L) or LTR (MMP2, CDKN2A, RUNX2, and RUNX3) retroelements and stomach-specific TFF3 gene were analyzed using radioisotope-labeled methylation-specific polymerase chain reaction.
RESULTS
Mean ages of H. pylori-positive controls with mild, moderate, and severe atrophy were 51, 54, and 65 years and those of H. pylori-associated TFF3 overmethylation at the three atrophic levels (51, 58, and 63 years) tended to be periodic. Alu-adjacent overmethylation (50 years) was earlier than TFF3 overmethylation (58 years) in H. pylori-positive controls with moderate atrophy. Cancer patients with moderate atrophy showed late Alu-adjacent (58 years) overmethylation and frequent LTR-adjacent overmethylation. LTR-adjacent overmethylation was frequent in cancer (66 years) and dysplasia (68 years) patients with severe atrophy.
CONCLUSION
Atrophic progression is associated with gastric cancer at moderate level by impeding the initiation of Alu-adjacent methylation. LTR-adjacent methylation is increased in cancer patients and subsequently in dysplasia patients.

Keyword

Helicobacter pylori; DNA methylation; Stomach neoplasms; Atrophic gastritis

MeSH Terms

Atrophy*
DNA Methylation
Epigenomics
Gastric Mucosa
Gastritis, Atrophic
Genes, Essential*
Helicobacter pylori
Housekeeping*
Humans
Methylation*
Polymerase Chain Reaction
Retroelements
Stomach Neoplasms*
Retroelements

Figure

• 

  Fig. 1. Comparison of numbers of overmethylated CpG-islandgenes among four subject groups: Helicobacter pylori–positive controls, H. pylori–negative controls, patients with gastric cancer, and patients with dysplasia. Numbers of overmethylated genes were separately calculated for CpG-island housekeeping genes adjacent to Alu retroelements (CDH1, ARRDC4, PPARG, and TRAPPC2L) or LTR retroelements (MMP2, CDKN2A, RUNX2, and RUNX3). (A) Numbers of overmethylated genes are represented as mean±standard error of mean (SEM) (*p< 0.05 compared to H. pylori–positive controls based on one-way ANOVA with Tukey’s post-hoc tests). (B) Numbers of overmethylated genes were further analyzed according to the level of gastric mucosal atrophy. They are presented as mean±SEM (*p< 0.05 compared to mild atrophic cases based on one-way ANOVA with Tukey’s post-hoc tests).

• 

  Fig. 2. Analysis of age-related methylation in Helicobacter pylori–positive controls. (A) Age-related methylation changes of TFF2, TFF3, and APC were evaluated in the antrum by mean level of methylation estimated by 10-level classification. Open and closed vertical arrows indicate low trough and high peak respectively. (B) Age-related methylation of antrum-specific TFF3 and body-specific GHRL was evaluated in the antrum and body by the frequency of overmethylation. Age-related methylation curves were plotted as a function of age-dependent APC-methylation level. (C) Age-related methylation of TFF3 in the antrum was analyzed according to three levels of gastric mucosal atrophy.

• 

  Fig. 3. Overmethylation age of Alu-adjacent genes, LTR-adjacent genes, atrophy-sensitive TFF3, and age-dependent APC in four subject groups. Each of the four subject groups was subgrouped according to three atrophic levels. (A, B) Mean age of subjects showing overmethylated Alu- and LTR-adjacent genes in the gastricbody and overmethylated TFF3 and APC genes in the gastric antrum were analyzed. (A) Mean ages were compared between cases with Alu- or LTR-adjacent overmethylation and with TFF3 overmethylation. (B) Mean ages were compared between cases with Alu- or LTR-adjacent overmethylation with and without TFF3 overmethylation. Error bars indicate standard error of mean. *p< 0.05 by Student’s t test. (C) Changes in proportions of Alu-adjacent overmethylation during the progression of gastric atrophy. Cases of Alu-adjacent overmethylation with and without antrum-site TFF3 overmethylation were evaluated for both gastric antrum and body sites. The vertical length of bars represents the proportion (%) of overmethylation cases according to three atrophic levels in each subject group. A horizontal line across the proportion bar indicates the mean age of each methylation group.

Reference

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