Korean J Gastroenterol.  2015 Aug;66(2):75-84. 10.4166/kjg.2015.66.2.75.

RUNX3 Methylation, Loss of RUNX3 Expression and Clinicopathologic Findings according to Helicobacter pylori CagA in Gastric Carcinoma

• Na YJ
• Shim KN
• Joo YH
• Kim SE
• Jung HK
• Jung SA
• Cho MS

• KMID: 2373318
• DOI: http://doi.org/10.4166/kjg.2015.66.2.75

Abstract

BACKGROUND/AIMS
Helicobacter pylori cytotoxin-associated gene A (CagA) has been suggested to be involved in the inactivation of Runt-related transcription factor 3 (RUNX3), a known gastric carcinoma tumor suppressor gene. It remains unclear how H. pylori CagA initiates or maintains RUNX3 promoter methylation and inactivates its protein expression in gastric carcinoma.
METHODS
RUNX3 promoter methylation status, RUNX3 expression, and H. pylori CagA were investigated in 76 sample pairs of gastric carcinoma tissue. The patients' medical records were reviewed. The association between RUNX3 methylation or loss of RUNX3 expression and clinicopathologic variables according to H. pylori CagA status were investigated.
RESULTS
In gastric carcinoma patients with H. pylori CagA-positive infection, RUNX3 methylation did not show association with lymphatic invasion, venous invasion, and TNM stages. However RUNX3 methylation was observed more frequently in poorly differentiated adenocarcinoma and signet ring cell carcinoma (77.8% vs. 20.0%, p=0.023) in early stage. In gastric carcinoma patients with H. pylori CagA-positive infection, loss of RUNX3 expression did not show association with lymphatic invasion, venous invasion, and TNM stages. However loss of RUNX3 expression was observed more frequently in early gastric carcinoma than in advanced gastric carcinoma (84.2% vs. 75.0%, p=0.51), but this difference was not significant.
CONCLUSIONS
In gastric carcinoma patients with H. pylori CagA-positive infection, RUNX3 methylation or loss of RUNX3 expression did not show correlation with lymphovascular invasion and TNM stages. In early gastric carcinoma patients with H. pylori CagA-positive infection, RUNX3 methylation was observed more in poorly differentiated adenocarcinoma and signet ring cell carcinoma.

Keyword

Helicobacter pylori; CagA; RUNX3; Methylation; Carcinoma

MeSH Terms

Adult
Aged
Aged, 80 and over
Antigens, Bacterial/*metabolism
Bacterial Proteins/*metabolism
Cell Line, Tumor
Core Binding Factor Alpha 3 Subunit/genetics/*metabolism
Female
*Gene Expression Regulation, Neoplastic
Helicobacter Infections/complications/microbiology
Helicobacter pylori/isolation & purification/*metabolism
Humans
Immunohistochemistry
Lymphatic Metastasis
Male
Methylation
Middle Aged
Neoplasm Staging
Promoter Regions, Genetic
Retrospective Studies
Stomach Neoplasms/complications/microbiology/*pathology
Antigens, Bacterial
Bacterial Proteins
Core Binding Factor Alpha 3 Subunit

Figure

• 

  Fig. 1. Representative results of methylation-specific PCR for RUNX3 and PCR for CagA in human normal and gastric cancer tissue samples. (A) Methylation specific PCR for RUNX3. Positive control, negative control, molecular weighted marker for RUNX3 and un-methylated bands (U) were seen in cancer tissue. (B) PCR for CagA. Presence of band in normal gastric mucosa or gastric carcinoma tissue was considered positive for Helicobacter pylori CagA.

• 

  Fig. 2. Immunohistochemical stain for RUNX3 protein. (A) Adjacent mucosa surrounded by tumor showed cytoplasmic positive reaction in goblet cells (black arrowhead). The lymphocytes in lamina propria showed a nuclear positive reaction of lymphocytes (blue arrowhead) (×200). (B) The tumor cells showed a negative reaction (×200). (C) The tumor cells showed a strong brownish nuclear positive reaction (×200). (D) The tumor cells showed no nuclear positive reaction but a cytoplasmic positive reaction (cytoplasmic mislocalization) (black arrowheads) (×400).

Reference

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