Adolescent ; Adult ; Aged ; Carcinoma ; metabolism ; pathology ; Core Binding Factor Alpha 3 Subunit ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Protein Array Analysis ; methods ; Young Adult

PURPOSE: We investigated the methylation pattern of RUNX3 to determine whether aberrant methylation events are risk factors for bladder tumor development, recurrence, and progression. MATERIALS AND METHODS: A hospital based, case-control investigation was carried out in 124 bladder tumor specimens and 20 normal bladder mucosae. The methylation pattern of RUNX3 was determined using MS- PCR and direct DNA sequencing. RUNX3 mRNA expression levels were assessed by RT-PCR. RESULTS: All normal bladder mucosae were unmethylated. On the other hand, bladder tumor tissues showed either unmethylated, methylated, or both unmethylated and methylated patterns. Methylation of RUNX3 promoter region was significant as a risk factor for bladder tumor development (p<0.01, OR=107.55, 95% CI=6.33-1827.39). The methylation type was more frequent in invasive tumors, compared with superficial bladder tumors (p=0.01, OR=2.95, 95% CI=1.16-7.47). Both recurrence (p=0.02, OR= 3.70, 95% CI=1.19-11.46) and subsequent tumor progression were significantly associated with the methylation of the RUNX3 promoter region (p=0.01, OR=5.63, 95% CI=1.23-25.82). The mutations in 3 cases out of 23 bladder tumor tissues were observed in contrast with normal bladder mucosae. CONCLUSIONS: These results suggest not only the possibility that RUNX3 is a tumor suppressor gene responsible for bladder tumor, but also that it may be a useful prognostic marker for bladder tumor recurrence and progression.
Case-Control Studies ; Core Binding Factor Alpha 3 Subunit ; Genes, Tumor Suppressor ; Hand ; Methylation ; Mucous Membrane ; Point Mutation ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Recurrence ; Risk Factors ; RNA, Messenger ; Sequence Analysis, DNA ; Urinary Bladder Neoplasms* ; Urinary Bladder*

BACKGROUND/AIMS: This study was performed to determine the association between RUNX3 expression and Helicobacter pylori infection in premalignant gastric lesions. METHODS: We examined 107 patients with gastric epithelial dysplasia who had undergone endoscopic mucosal resection or submucosal dissection. All tissue samples were evaluated by RUNX3 staining and subclassified by immunophenotype. H. pylori infection in dysplastic lesions and the normal surrounding tissue was examined by silver staining, and cagA status was assessed by polymerase chain reaction. RESULTS: The loss of RUNX3 expression was observed in 62 cases (57.9%), and an association with H. pylori infection was found in 54 cases (50.5%). The infection rate with the cagA-positive H. pylori strain was 63.0%. In RUNX3-negative lesions, the rate of H. pylori infection (p=0.03) and the frequency of category 4 lesions (according to the revised Vienna classification) were high (p=0.02). In addition, the gastric mucin phenotype was predominant. In RUNX3-negative category 4 lesions, the rate of cagA-positive H. pylori infection rate was high but not significantly increased (p=0.08). CONCLUSIONS: Infection with H. pylori is associated with inactivation of RUNX3 in early gastric carcinogenesis. This mechanism was prominent in gastric cancer with a gastric mucin phenotype.
Adenoma/*chemistry ; Aged ; Antigens, Bacterial/genetics ; Bacterial Proteins/genetics ; Carcinoma/*chemistry ; Cell Transformation, Neoplastic ; Core Binding Factor Alpha 3 Subunit/*analysis ; Female ; Gastric Mucosa/*chemistry/pathology ; Helicobacter Infections/*metabolism ; Helicobacter pylori/*genetics ; Humans ; Male ; Middle Aged ; Mucin 5AC/analysis ; Mucin-2/analysis ; Mucin-6/analysis ; Neprilysin/analysis ; Phenotype ; Precancerous Conditions/*chemistry/pathology ; Stomach Neoplasms/*chemistry

OBJECTIVETo investigate the role of Runx3 gene CpG island methylation in the development of human gastric carcinoma.

METHODSA total of 150 tumor specimens from patients with gastric carcinoma and 50 normal tissue specimens were selected. Methylation specific PCR (MSP) and pyrosequencing (PS) were used to detect the methylation status of Runx3 gene promoter.

RESULTSCompared to normal tissue samples, a significant increase of CpG island methylation status of Runx3 gene was observed in gastric carcinomas (MSP: 67.3% vs. 40.0%, P = 0.002; PS: 76.0% vs. 30.0%, P < 0.01). Runx3 gene methylation was only related to tumor size (P < 0.05) based on MSP analysis. PS test however showed that the extent of methylation of Runx3 gene was related to the tumor size (P = 0.004), Lauren's classification (P = 0.043), depth of invasion (P < 0.01), lymph node metastasis (P = 0.021) and TNM staging (P = 0.045).

CONCLUSIONSMethylation status of Runx3 gene detectable by PS is closely correlated with clinicopathological parameters of gastric carcinoma, including tumor size, Lauren's classification, depth of invasion, lymph node metastasis and TNM staging. PS is more sensitive than MSP in the detection of Runx3 gene methylation, which may serve as an important marker for early diagnosis and treatment of gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; CpG Islands ; genetics ; DNA Methylation ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tumor Burden ; Young Adult