Relative expression of miR-196a2 and targets in cancer tissues compared to control.

<p>(A-D) Box plots represent median (quartiles) of gene fold change stratified by cancer type, calculated using LIVAC method. EC, esophageal cancer; GC, gastric carcinoma; SIC, small intestine cancer; CRC, colorectal carcinoma; red dotted lines are equivalent for relative expression at 1.0 of controls. Mann-Whitney U test was used. Statistical significance at <i>P</i><0.05. (E) Annexin A1 protein expression assessed by immunohistochemistry (IHC).</p

Evaluation of miRNA-196a2 and apoptosis-related target genes: <i>ANXA1</i>, <i>DFFA</i> and <i>PDCD4</i> expression in gastrointestinal cancer patients: A pilot study

<div><p>Previous reports have suggested the significant association of miRNAs aberrant expression with tumor initiation, progression and metastasis in cancer, including gastrointestinal (GI) cancers. The current preliminary study aimed to evaluate the relative expression levels of miR-196a2 and three of its selected apoptosis-related targets; <i>ANXA1</i>, <i>DFFA</i> and <i>PDCD4</i> in a sample of GI cancer patients. Quantitative real-time PCR for miR-196a2 and its selected mRNA targets, as well as immunohistochemical assay for annexin A1 protein expression were detected in 58 tissues with different GI cancer samples. In addition, correlation with the clinicopathological features and <i>in silico</i> network analysis of the selected molecular markers were analyzed. Stratified analyses by cancer site revealed elevated levels of miR-196a2 and low expression of the selected target genes. Annexin protein expression was positively correlated with its gene expression profile. In colorectal cancer, miR-196a over-expression was negatively correlated with annexin A1 protein expression (r = -0.738, <i>p</i> < 0.001), and both were indicators of unfavorable prognosis in terms of poor differentiation, larger tumor size, and advanced clinical stage. Taken together, aberrant expression of miR-196a2 and the selected apoptosis-related biomarkers might be involved in GI cancer development and progression and could have potential diagnostic and prognostic roles in these types of cancer; particularly colorectal cancer, provided the results experimentally validated and confirmed in larger multi-center studies.</p></div

Summary of the role of miR-196a2 and the selected targets in gastrointestinal tumors of the current study.

<p>As a diagnostic (A) and prognostic (B) tumor markers in esophageal, gastric, small intestine, and colorectal cancer.</p

Summary of previously published validation studies for the current miR-196a2 putative targets.

<p>Summary of previously published validation studies for the current miR-196a2 putative targets.</p

Enrichment functional analysis of hsa-miR-196a2 gene targets.

<p>Fisher's Exact test (hypergeometric distribution) was applied as an enrichment analysis method (p = 5.868485e-07). Black arrow highlighted one of the gene ontology (Cell Death) of the gene set [Data source: mirPath v.3].</p

Immunohistochemical analysis of annexin A1 protein expression in GI cancer.

<p>(1) Normal esophageal mucosa normally express annexin A1 nuclear and cytoplasmic (x 200). Esophageal GII squamous expressed annexin A1 (nuclear) with score 4 (2x200); Esophageal adenocarcinoma grade II expressed annexin A1 (nuclear) with score 3 (3x200); while grade III esophageal adenocarcinoma with score 6 (4 x100). Gastric adenocarcinoma grade I expressed annexin A1 with score 9 (5x100); grade II expressed it with score 3 (6 x200); while Undifferentiated gastric carcinoma, and gastric signet ring carcinoma (8) didn’t express annexin A1 (score 0) (7 x200) (8 x200) respectively; Small intestinal villous adenoma express annexin A1 nuclear and cytoplasmic with score 3 (9 x200); Small intestinal adenocarcinoma grade I express annexin A1 cytoplasmic and nuclear with score 6 (10 X200); while lesser expression of annexin A1 score 3 was detected in small intestinal adenocarcinoma grade II (11 x200). Normal colonic mucosa didn’t express annexin A1 (score 0) (12 X100); Colonic adenocarcinoma grade I expressed annexin A1 with score 9 (13x100); Colonic adenocarcinoma grade I infiltrating subserosal fat expressed annexin A1with score 9 (14x100); Colonic adenocarcinoma grade II expressed annexin with score 4 (15x200); (16) Colonic adenocarcinoma grade II infiltrating muscle and didn’t express annexin A1 score 0 (annexin 16 x200); Colonic adenocarcinoma grade III and colonic mucinous carcinomas expressed annexin A1 in less than 5% of tumor score 0 (17 x200); (18 x100), respectively.</p

Association of MicroRNA-196a2 Variant with Response to Short-Acting β2-Agonist in COPD: An Egyptian Pilot Study

<div><p>Chronic obstructive pulmonary disease (COPD) is a multifactorial chronic respiratory disease, characterized by an obstructive pattern. Understanding the genetic predisposition of COPD is essential to develop personalized treatment regimens. MicroRNAs (miRNAs) are small, endogenous, non-coding RNAs that modulate the expression levels of specific proteins based on sequence complementarity with their target mRNA molecules. Emerging evidences demonstrated the potential use of miRNAs as a disease biomarker. This pilot study aimed to investigate the association of the MIR-196a2 rs11614913 (C/T) polymorphism with COPD susceptibility, the clinical outcome and bronchodilator response to short-acting β₂-agonist. Genotyping of rs11614913 polymorphism was determined in 108 COPD male patients and 116 unrelated controls using real-time polymerase chain reaction technology. <i>In silico</i> target prediction and network core analysis were performed. COPD patients did not show significant differences in the genotype distribution (<i>p</i> = 0.415) and allele frequencies (<i>p</i> = 0.306) of the studied miRNA when compared with controls. There were also no associations with GOLD stage, dyspnea grade, disease exacerbations, COPD assessment test for estimating impact on health status score, or the frequency of intensive care unit admission. However, COPD patients with CC genotype corresponded to the smallest bronchodilator response after Salbutamol inhalation, the heterozygotes (CT) had an intermediate response, while those with the TT genotype showed the highest response (<i>p</i> < 0.001). In conclusion MIR-196a2 rs11614913 polymorphism is associated with the bronchodilator response of COPD in our sample of the Egyptian population, generating hypothesis of the potential use of MIR-196a2 variant as a pharmacogenetic marker for COPD.</p></div

Clinical and functional characteristics of COPD patients and controls.

<p>Clinical and functional characteristics of COPD patients and controls.</p

STRING analysis of miR-196a2 targets.

<p>(A) Protein-protein interaction network. Network nodes represent proteins, target genes are encircled in red, edges represent protein-protein associations, colored according to action types between proteins. The network of the three target genes (<i>ANXA1</i>, <i>PDCD4</i>, and <i>DFFA</i>) composed of 18 nodes and 57 edges with clustering coefficient of 0.803. (B) Enrichment analysis showing gene ontology of biological processes. Red nodes are the target genes, yellow box specific for <i>ANXA1</i>, blue for <i>PDCD4</i>, orange for <i>DFFA</i>, green; function shared by <i>ANXA1</i> and <i>PDCD4</i>, and pink for biological processes implicated by the three genes.</p

Predicted miRNA: Gene interactions.

<p>The predicted miRNA (red color)::mRNA (blue color) duplex. The putative target sites predicted by miRanda are scored for likelihood of mRNA down-regulation using mirSVR, a regression model that is trained on sequence and contextual features of the predicted miRNA::mRNA duplex. The colon (:) represents the location of a G/U wobble base pairing [Data source: microRNA.org].</p