No association between the SNP rs1625579 in miR-137 gene and schizophrenia in Iranian population

Background: Schizophrenia is a common and severe neuropsychiatric disorder with symptoms such as hallucination, delusion and mental disease. Among candidate genes for schizophrenia, miR-137 gene has been recently suggested to contribute to schizophrenia by genome-wide association study (GWAS) and a single nucleotide polymorphism (SNP) rs1625579 (G>T) as a presumed risk allele within an intron of miR-137 gene has been contributed by schizophrenia. Since mir-137 is a considerable gene in the performance of neural systems, the present study dealt with the association between SNP rs1625579 in miR-137 gene and schizophrenia in Iranian patients. Aim of study: This study aimed to evaluate the association between SNP rs1625579 in miR-137 gene and schizophrenia in Iranian patients. Methods: Hoping to identify a single-nucleotide polymorphism as a possible locus for schizophrenia, we carried out this case-control study on 80 blood samples collected from individuals suffering from schizophrenia and 48 healthy controls. DNA was extracted from the samples, and the frequency of the polymorphisms was analyzed using ARMS-PCR method. Finally, the products were detected on 1.5% agarose gel electrophoresis. Results: The analysis on the data showed that 43.75% of the patients and 37.5% of the controls were mutant homozygous and 56.25% of the patients and 62.5% of controls were heterozygous. In addition, 0.0% of the patients and 0.0% of the controls were normal homozygous. Both the genotype (p = .48 > .05) and allele (p = .5 > .05) distribution of the rs1625579 SNP has no significant difference between patients and controls. Conclusion: There was no significant relationship between rs1625579 and the incidence of schizophrenia. To the best of our knowledge, this is first study in Iran that assesses the frequency of the polymorphism among Iranian patients. However, further studies with more samples are necessary. Keywords: MIR137 gene, Rs1625579, Schizophrenia, Polymorphism, Ira

Upregulation of miR-371-373 cluster, a human embryonic stem cell specific microRNA cluster, in esophageal squamous cell carcinoma

Aims: Esophageal squamous cell carcinoma (ESCC) is the most common subtype of esophageal cancer in Iran. MicroRNAs (miRNAs) are a class of noncoding RNAs that are found to be involved in different processes and can play a role in tumorigenesis and result in cancer. MiR-371, miR-372, and miR-373 are a gene cluster that is located in the region of the human chromosome of 19q13.4. They are specifically expressed in human embryonic stem cells (ESCs) and involved in the maintenance of the stemness features through regulating the expression of certain key genes and signaling pathways. The present study investigated the potential expression of miR-371–373 cluster in tumor and nontumor tissues of ESCC. Materials and Methods: The expression level of miR-371–373 cluster was analyzed in paraffin-embedded tissues of tumor and tumor margin in 36 patients with ESCC. Total RNA was isolated and the miR-371–373 clusters were quantified with quantitative real-time-polymerase chain reaction expression analysis. Computed tomography analysis (2–ΔΔCT) and t-test were used to determine the relationship between the characteristics of the tumor and nontumor tissues. Statistically, P value of <0.05 were considered significant. Data analysis was performed using SPSS 16. Results: We provided miR-371, miR-372, and miR-373 upregulation evidence significantly with 14.36, 26.9, and 21.1-fold in esophageal cancer cells compared with their adjacent normal cells (P < 0.05), respectively. In addition, evaluation of these genes expression in various grades didn't show a significant difference. Conclusion: Our findings support the hypothesis that these miRNAs might play a role in tumorigenesis in esophageal cancer

<em>In vitro</em> cytotoxic and apoptotic activities of <em>Allium paradoxum</em> (M. Bieb.) G. Don extract on human breast cancer cell line

247-254Researchers from all pharmaceutical fields are trying to find new drugs from natural origin with less toxicity. In northern Hyrcanian forests Iran, Allium paradoxum (M. Bieb.) G. Don has traditionally used as food and vegetable. Previously studies reports, this plant has a medicinal potential for anti-oxidant and anti-hemolytic activities. In this regard, we evaluated the anti-tumor activity of hydroalcoholic extract of A. paradoxum (M. Bieb.) G. Don in different concentrations on human breast cancer cell line (MCF-7). MTT assay was performed with MCF-7 cancer cell line and also evaluation of apoptotic effect, Bax and Bcl-2 expression in MCF7 cells were analyzed by real time RT-PCR. The results showed that the A. paradoxum (M. Bieb.) G. Don extracts decrease the viability of MCF-7 cell line in a dose-dependent manner and the most effective concentration of this extracts after 24 h treatment was 100 μM. Apoptosis induction was confirmed by fluorescence microscopy and plant extracts display a pro-apoptotic effect by down-regulated and up-regulated the expression of Bcl-2 and BAX in tumor cells, respectively. In conclusion, the study was confirmed pro-apoptotic and cytotoxicity effect of A. paradoxum (M. Bieb.) G. Don extract against MCF-7 cell lines. Based on being natural, low cost, accessibility, and noteworthy advantages of this product, it seems that A. paradoxum (M. Bieb.) G. Don has a potential source for isolation of novel anticancer agents for a drug

MiR-21 expression is mainly confined to the tumor stroma.

<p>A) <i>In situ</i> hybridization in FFPE samples of esophageal cancer localized miR-21 expression (blue signals) in cancer associated fibroblasts of the tumor stroma, but not in the tumor cells. Slides were counterstained with nuclear fast red. B) The adjacent normal squamous part on the same slide did not show miR-21 expression neither in the stroma nor in the squamous cells; C) Nuclear staining of U6 snRNA was used as an internal control; D) Negative control without probe. Bottom-left inserts show a 2 times bigger magnification of each image; E) Samples with high stromal component showed significantly higher levels of miR-21 expression than samples with a low stromal content. (P value  = 0.04 with unpaired T-test with Welch' s correction).</p

Histopathological criteria of the analyzed SCC patients.

<p>Histopathological criteria of the analyzed SCC patients.</p

Differential expression of miR-21 in 42 FFPE tumor samples in comparison with adjacent non-tumoral tissue.

<p>Quantitative RT-PCR analysis on FFPE samples of esophageal SCC patients shows higher levels of miR-21 in cancerous tissue as compared to the adjacent non-cancerous counterpart (P = 0.0007). MicroRNA levels are normalized to 5S rRNA. Values are presented as means ± standard deviation. The P value was determined with a 2-tailed Student' s t-test.</p

Effect of co-culturing normal fibroblasts with esophageal cancer cell lines on miR-21 expression levels.

<p>A) A schematic view of the co-culture system; B) MiR-21 expression in KYSE-30 cells and normal fibroblasts (HGF-1) after juxtaposition in a co-culture system; C) MiR-21 expression levels in KYSE-30 and HGF-1 cells after treatment with CM of HGF-1 and KYSE-30 cells, respectively; D) MiR-21 expression levels in FLO-1 adenocarcinoma and HGF-1 fibroblast cells after juxtaposition in a co-culture system; E) MiR-21 expression levels in FLO-1 and HGF-1 cells after treatment with CM of HGF-1 and FLO-1 cells, respectively. Data were normalized to expression after 24 h of co-culture. Each experiment was performed at least 2 times in triplicates.</p

Cell migration and invasion properties of KYSE-30 cells are increased upon co-culture with HGF-1.

<p>A) KYSE-30 cells juxtaposed with HGF-1 fibroblasts showed a significantly higher potential to migrate (P<0.0001) or invade (P = 0.0001) through the coated 8.0 μm pore-sized membrane as compared to the KYSE-30 cells not grown co-culture with normal fibroblasts. Images of representative microscopic pictures are shown on the left; B) 3 day old conditioned media from HGF-1 fibroblasts could significantly induce migration and invasion of KYSE-30 cells (P<0.0001 for both experiments). Images of representative microscopic pictures are shown on the left.</p

Expression analysis of 6 fibroblastic markers in normal fibroblasts co-cultured with esophageal cancer cell lines KYSE-30 (A), FLO-1 (B) and OE-33 (C).

<p><i>COL4A1</i> expression in fibroblasts was significantly decreased after 2 days of incubation with KYSE-30 (P = 0.03) and OE-33 (P = 0.04) cells and after 3 days of incubation with FLO-1 cells (P = 0.01). <i>TIMP3</i> expression was <i>s</i>ignificantly decreased after 2 and 3 days of incubation with KYSE-30 (P = 0.0002 and P>0.0001, respectively), after 2 and 3 days of incubation with FLO-1 cells (P = 0.0013 and P = 0.0006, respectively) and after 2 and 3 days of incubation with OE-33 cells (P = 0.0017 and P = 0.0004, respectively) when compared to the measurement 24 h after the start of the co-culture.</p