Associations between methylation of EMT related genes and subtypes of UC.

<p>Note: Data are expressed as the mean ± SE. n, number of samples.</p><p>Two and three groups were compared using the t-test and Kruskal-Wallis test, respectively.</p>¶<p>? #, , & and *: Data are missing for one, three, one, and one cases, respectively.

: <i>CDH1</i>, <i>p</i> = 0.09, &: <i>CDX1</i>, <i>p</i> = 0.04, <i>miR-1247</i>, <i>p</i> = 0.006, <i>CDH1</i>, <i>p</i> = 0.08.*: <i>miR-1247</i>, <i>p</i> = 0.06.</p><p>Associations between methylation of EMT related genes and subtypes of UC.</p

Methylation of <i>miR-1247</i> (left), <i>CDH1</i> (center) and mean Z score of the two genes (right) in relation to the Mayo endoscopic subscore.

<p>The statistical analysis was performed using one way ANOVA.</p

Unsupervised hierarchical clustering analysis using paired inflammatory and non-inflammatory colonic mucosa derived from ten patients.

<p>Black boxes, inflammatory rectal mucosa (R); grey boxes, normal proximal mucosa (N); Samples of the same ID number were obtained from the same patients.</p

Primer sequences used in pyrosequencing.

<p>U =  biotin labeled universal primer tag: 5′-biotin-GGGACACCGCTGATCGTTTA.</p><p>Primer sequences used in pyrosequencing.</p

Association between methylation status of EMT-related genes and global hypomethylation assessed by methylation of the <i>LINE1</i> repetitive element.

<p>Statistical analysis was performed using the Spearman correlation analysis.</p

Methylation of <i>miR-1247</i> (left), <i>CDH1</i> (center) and mean Z score of the two genes (right) in relation to severe clinical phenotype of UC.

<p>The statistical analysis was performed using Student's t-Test.</p

Methylation of <i>CDH1</i> (left), <i>CDH13</i> (center) and mean Z score of the two genes (right) in relation to the age and duration of disease.

<p>Statistical analysis was performed using a Spearman correlation analysis.</p

Change in DNA Methylation Patterns of <i>SLC6A4</i> Gene in the Gastric Mucosa in Functional Dyspepsia

<div><p>Background</p><p>The neurochemical serotonin (5-HT) is an important signaling molecule in the gastrointestinal motor and sensory functions. A key regulator of 5-HT levels is the transmembrane serotonin transporter (5-HTT; SLC6A4) that governs the reuptake of 5-HT. Recent studies have indicated 5-HTT expression may be regulated by epigenetic mechanisms. We investigated DNA methylation status of <i>SLC6A4</i> gene in the gastric mucosa from functional dyspepsia (FD) because of their potential role in dyspeptic symptoms.</p><p>Methods</p><p>Endoscopic gastric biopsies were obtained from 78 subjects with no upper abdominal symptoms and 79 patients with FD. Bisulfite Pyrosequencing was carried out to determine the methylation status of promoter CpG islands (PCGIs), promoter non-CpG islands (PNCGIs) and gene body non-CpG islands (NPNCGIs) in the <i>SLC6A4</i> gene. Gene expression was examined by real-time PCR.</p><p>Results</p><p>In overall, methylation level of PCGIs was significantly lower in FD compared to control subjects (<i>p</i> = 0.04). On the other hand, methylation level of NPNCGIs was significantly higher in FD compared to control subjects (<i>p</i> = 0.03). Lower methylation level in PNCGIs was highlighted in the patients with PDS (<i>p</i> = 0.01), while higher methylation level in NPNCGIs was more prominent in the patients with EPS (<i>p</i> = 0.017). Methylation levels of PCGIs and PNCGIs were inversely correlated, while methylation levels of NPNCGIs was positively correlated with SLC6A4 mRNA levels in FD patients.</p><p>Conclusions</p><p>Our data suggest that change in DNA methylation pattern of SLC6A4 in the gastric mucosa may have a role for developing FD. A role of epigenetics for developing FD needs to be further evaluated.</p></div

Representative bisulfite cloning sequencing of the <i>SLC6A4</i> PCGI, PNCGI (-36∼+499) and NPNCGI (+22987∼+23308) in FD and control.

<p>Open and filled circles represent unmethylated and methylated CpG sites, respectively. Transcriptional start site is by indicated by an arrow on the top. CpG islands (CGI) are indicated as green bar on the top.</p

Structure of <i>SLC6A4</i> gene and the PCGI, PNCGI and NPNCGI regions analyzed by bisulfite pyrosequencing (black bars) and bisulfite cloning sequencing (grey bars).

<p>CpG islands represent green bar (a). A representative pyrogram from a clinical sample (b).</p