5-Azacytidine Enhancing Expression of E-cadherin in Adenocarcinoma Cell Line

Introduction: In this study, we assessed the expression of E-cadherin in HT29 cell line treated with 5-Azacytidine and colorectal cancer patient in an Iranian population. E-cadherin expression promotes metastasis and prognosis of colorectal cancer (CRC). 5-Azacytidine, a DNA methyl transferase inhibitor, is a clinically used epigenetic drug for treatment of cancer including colorectal cancer, leading to genes activation involved in tumor suppression, especially E-cadherin. Materials and Methods: HT29 cell line treated with 5-Azacitidine and 40 polyps, 20 tumors and 40 adjacent normal tissues samples were enrolled in this study. Using the real-time PCR method, the expression levels of E-cadherin were examined in treated cell line and colorectal cancer tissue. Results: This study proves that 5-Azacytidine induces over expression of E-cadherin in adenocarcinoma cell line, while the expression levels of E-cadherin were not different in tumor and polyp than adjacent normal tissue. Conclusion: To conclude, 5-Azacytidine induces re-expression of E-cadherin in adenocarcinoma cell line. Thus, 5-Azacytidine as demethylation drug activated tumor suppressor gene as E-cadherin

Comparison of Insulin Expression Levels in White Blood Cells of infants with and without Family History of Type II Diabetes

Background: Type II diabetes is known as one of the most important, prevalent, and expensive diseases of mankind. Late diagnosis and subsequent delayed initiation of treatment or surveillance of patients create a variety of problems for affected individuals. This has raised increasing concerns for public health authorities throughout the world. In the current study, we aimed to find a new approach for early identification of high-risk individuals at initial months of their life. This allows us to take preventive measures as early as possible.Materials and Methods: In our study, 102 infants - from one to six months - were selected and placed in two case and control groups. The case group contained 52 babies with at least one of their parents identified as a type II diabetic patient. The control group comprised 50 babies with no family history of type II diabetes in paternal and maternal first-degree relatives. Afterwards, the expression level of insulin gene was analyzed in white blood cells of both groups. Information related to infants - referred to outpatient and inpatient wards of three main pediatric hospitals placed in Tehran - and their parents were collected through questionnaires within a two-year period. The study inclusion criteria for infants were confirmed type II diabetes in at least one of their parents, the absence of any metabolic disorder, and the absence of any disturbing vital signs. After drawing 2 ml of babies’ peripheral blood, total RNA of white blood cells (WBC) was extracted, and used for cDNA synthesis. Real-Time PCR was then applied to quantitatively evaluate the expression levels of insulin gene. The results of Real-Time PCR were statistically analyzed by non-parametric tests of Mann-Whitney and Kruskal-Wallis.Results: The expression of insulin gene was observed in white blood cells of all samples. However, there was a significant difference in expression levels between case and control groups (p<0.05). There was a statistically significant difference in mean levels of gene expression among babies with diabetic mother, and healthy groups (RQ=0.5, P-value=0.002), but this value wasn’t significant for babies with diabetic father (RQ=0.78, P>0.05).Conclusion: Numerous genes contribute to the development of diabetes and novel disease-causing genes are increasingly being discovered. Identification of disease-prone individuals through examining merely one underlying gene is complicated and challenging. Interestingly, all of these abnormally functioning genes finally manifest themselves in the altered expression levels of insulin gene. The expression status of insulin gene in WBCs could be suggested as a useful approach for identification of individuals at high risk for developing diabetes. This paves the way for taking appropriate measures at infancy period in order to prevent the disease as well as inhibit its various side effects in the following years of patient’s life

Cytogenetic Abnormalities and Y Chromosome Microdeletions in Azoospermic and Oligospermic Infertile Males from West of Iran

     About 15% of couples have infertility problems, half of which are related to male factors. Cytogenetic and genetic disorders account for about 10% of the male infertility problems. The aim of this study was to determine the frequency and types of both cytogenetic abnormalities and AZF microdeletions of Y chromosome in idiopathic azoospermic and oligospermic infertile men in west of Iran. In this case-control study, a total of 108 infertile men including 62 azoospermic and 46 oligospermic men were studied for the cytogenetic and AZF microdeletions. Moreover, 90 fertile men served as a control group. Detailed clinical and laboratory examination was done for all participants. Karyotyping was done on peripheral blood lymphocytes to detect the cytogenetic abnormalities; likewise, multiplex-PCR method was performed to identify the presence of microdeletion in AZFa, AZFb or AZFc regions. Chromosomal abnormalities were detected in 6.5% (7/108) of cases, including two oligospermic men with balanced autosomal rearrangements, one oligospermic and four azoospermic men with Klinefelter syndrome. Y chromosome microdeletions were detected in 4.6% (5/108) of infertile men (AZFc: 3.7%, AZFbc: 0.9%). No AZFa deletion was detected in any of the patients. No chromosomal abnormality and Y chromosome microdeletion was detected in control group. The prevalence of chromosomal abnormalities and Y chromosome microdeletions shows the importance of genetic factors in male infertility. The analysis of karyotype and Y microdeletions in infertile men provide a proper understanding about the causes of infertility, the choice of the appropriate assisted reproduction technique and reducing the risk of transmission of these genetic defects to the future generation.

Autophagy ATG16L1 rs2241880 impacts the colorectal cancer risk: A case-control study

Background Despite many efforts to discover the important role of the autophagy process in the pathogenesis of colorectal cancer (CRC), the exact involved molecular mechanism still remains to be elucidated. Recently, a limited number of studies have been employed to discover the impact of autophagy genes' variants on the development and progression of CRC. Here, we evaluated the association between two single-nucleotide polymorphisms (SNPs) in the main components of the autophagy genes, ATG16L1 rs2241880, and ATG5 rs1475270, and the CRC risk in an Iranian population. Methods During this investigation, a total of 369 subjects, including 179 CRC patients and 190 non-cancer controls have been genotyped using Tetra-primer amplification refractory mutation system-polymerase chain reaction (TP-ARMS-PCR) method. Result The results demonstrated that the T allele of the ATG16L1 rs2241880 was significantly associated with the increased risk of CRC in the studied population (OR 1.64, 95% CI: 1.21-2.22, p = 0.0015). Moreover, ATG16L1 rs2241880 TT genotype increased the susceptibility to CRC (OR 3.31, 95% CI: 1.64-6.69, p = 0.0008). Furthermore, a significant association was observed under the recessive and dominant inheritance models (p = 0.0015 and p = 0.017, respectively). No statistically significant differences were found in the ATG5 rs1475270 alleles and genotypes between the cases and controls. Conclusion The results of the present study may be helpful concerning the risk stratification in CRC patients based on the genotyping approach of autophagy pathways and emphasize the need for further investigations among different populations and ethnicities to refine our conclusions

Bioinformatics prioritization of SNPs perturbing microRNA regulation of hematological malignancy-implicated genes.

The contribution of microRNAs (miRNAs) to cancer has been extensively investigated and it became obvious that a strict regulation of miRNA-mRNA regulatory network is crucial for safeguarding cell health. Apart from the direct impact of miRNA dysregulation in cancer pathogenesis, genetic variations in miRNAs are likely to disrupt miRNA-target interaction. Indeed, many evidences suggested that SNPs within miRNA regulome are associated with the development of different hematological malignancies. However, a full catalog of SNPs within miRNAs target sites of genes relevant to hematopoiesis and hematological malignancies is still lacking. Accordingly, we aimed to systematically identify and characterize such SNPs and provide a prioritized list of most potentially disrupting SNPs. Although in the present study we did not address the functional significance of these potential disturbing variants, we believe that our compiled results will be valuable for researchers interested in determining the role of target-SNPs in the development of hematological malignancies

Autophagy ATG16L1 rs2241880 impacts the colorectal cancer risk: A case-control study

Background: Despite many efforts to discover the important role of the autophagy process in the pathogenesis of colorectal cancer (CRC), the exact involved molecular mechanism still remains to be elucidated. Recently, a limited number of studies have been employed to discover the impact of autophagy genes’ variants on the development and progression of CRC. Here, we evaluated the association between two single-nucleotide polymorphisms (SNPs) in the main components of the autophagy genes, ATG16L1 rs2241880, and ATG5 rs1475270, and the CRC risk in an Iranian population. Methods: During this investigation, a total of 369 subjects, including 179 CRC patients and 190 non-cancer controls have been genotyped using Tetra-primer amplification refractory mutation system-polymerase chain reaction (TP-ARMS-PCR) method. Result: The results demonstrated that the T allele of the ATG16L1 rs2241880 was significantly associated with the increased risk of CRC in the studied population (OR 1.64, 95% CI: 1.21–2.22, p = 0.0015). Moreover, ATG16L1 rs2241880 TT genotype increased the susceptibility to CRC (OR 3.31, 95% CI: 1.64–6.69, p = 0.0008). Furthermore, a significant association was observed under the recessive and dominant inheritance models (p = 0.0015 and p = 0.017, respectively). No statistically significant differences were found in the ATG5 rs1475270 alleles and genotypes between the cases and controls. Conclusion: The results of the present study may be helpful concerning the risk stratification in CRC patients based on the genotyping approach of autophagy pathways and emphasize the need for further investigations among different populations and ethnicities to refine our conclusion

Analysis of Cytogenetic Abnormalities in Iranian Patients with Syndromic Autism Spectrum Disorder: A Case Series

Objective Autism spectrum disorder is a heterogeneous neuropsychiatric group of pervasive development disorder, which is mostly diagnosed through the intricate behavioral phenotype. According to strong genetic involvement, detecting the chromosome regions and the key genes linked to autism can help to elucidate its etiology. The present study aims to investigate the value of cytogenetic analysis in syndromic autism as well as to find an association between autism and chromosome abnormalities. Materials & Methods Thirty-six autism patients from 30 families, diagnosed clinically with DSM-5 criteria, were recruited. The syndromic patients who had additional clinical features involving development delay, attention deficit, hyperactivity disorder, seizure, language, and intellectual impairment were selected due to elevating the detection rate. Cytogenetics analysis was performed using GTG banding on the patients' cultured fibroblasts. Moreover, array-comparative genomic hybridization was also performed for a patient with a de novo and novel variant.   Results Karyotype analysis in 36 syndromic autism patients detected chromosomal abnormalities in two (5.6%) families, including 46,XY,dup(15)(q11.1q11.2) and 46,XX,ins(7)(q11.1q21.3)dn. In the latter, array-comparative genomic hybridization detected three abnormalities on chromosome 7, including deletion and insertion on both arms; 46,XX,del(7)(q21.11q21.3),dup(7)(p11.2p14.1p12.3)dn. Conclusion We reported a novel and de novo cytogenetic abnormality on chromosome 7 in an Iranian patient diagnosed with syndromic autism. However, the detection rate in syndromic autism was low which implies that it cannot be utilized as the only diagnostic procedure

Dysregulation of vitamin D synthesis pathway genes in colorectal cancer: A case-control study

Background: The cytochromes P450 are a superfamily of enzymes that control the synthesis of the biologically active form of vitamin D, 1,25-dihydroxyvitamin D3. These enzymes contribute to the formation of 1,25-dihydroxyvitamin D3, which starts with a 25-hydroxylation by CYP2R1 and CYP27A1 and a subsequent 1α-hydroxylation via CYP27B1. Methods: By using quantitative real-time polymerase chain reaction (qRT-PCR), we analyzed the expression ratio of CYP2R1, CYP27A1 and CYP27B1 genes within the vitamin D metabolic pathway in a total of 75 colorectal cancer (CRC) tissues compared to the adjacent tissues. Furthermore, we evaluated the association of CYP27B1 rs4646536 and CYP2R1 rs12794714 and rs10766196 polymorphisms with CRC risk in a total of 490 subjects, including 245 CRC patients and 245 non-cancer controls. The genotyping was performed using tetra-primer amplification refractory mutation system polymerase chain reaction (TP-ARMS�PCR) method. Results: The results indicated 2.3 and 2.7 upregulation of CYP2R1 and CYP27B1 genes in colorectal cancer tissues compared to the adjacent tissues, respectively. Rs12794714 AG genotype increased the risk of CRC (P =.03). Furthermore, a significant association was observed under the dominant inheritance model (P =.039). Conclusion: CYP2R1 and CYP27B1 genes were over-expressed in CRC samples compared to the adjacent control tissues. Furthermore, CYP2R1 rs12794714 variant was associated with the risk of CRC in the studied samples. CYP2R1 rs10766196 and CYP27B1 rs4646536 are not responsible for CYP2R1 and CYP27B1 genes expression alteration, respectively, but CYP2R1 rs12794714 polymorphism may be the reason of CYP2R1 upregulation and increased the risk of CRC

Dysregulation of vitamin D synthesis pathway genes in colorectal cancer: A case-control study

Background: The cytochromes P450 are a superfamily of enzymes that control the synthesis of the biologically active form of vitamin D, 1,25-dihydroxyvitamin D3. These enzymes contribute to the formation of 1,25-dihydroxyvitamin D3, which starts with a 25-hydroxylation by CYP2R1 and CYP27A1 and a subsequent 1α-hydroxylation via CYP27B1. Methods: By using quantitative real-time polymerase chain reaction (qRT-PCR), we analyzed the expression ratio of CYP2R1, CYP27A1 and CYP27B1 genes within the vitamin D metabolic pathway in a total of 75 colorectal cancer (CRC) tissues compared to the adjacent tissues. Furthermore, we evaluated the association of CYP27B1 rs4646536 and CYP2R1 rs12794714 and rs10766196 polymorphisms with CRC risk in a total of 490 subjects, including 245 CRC patients and 245 non-cancer controls. The genotyping was performed using tetra-primer amplification refractory mutation system polymerase chain reaction (TP-ARMS�PCR) method. Results: The results indicated 2.3 and 2.7 upregulation of CYP2R1 and CYP27B1 genes in colorectal cancer tissues compared to the adjacent tissues, respectively. Rs12794714 AG genotype increased the risk of CRC (P =.03). Furthermore, a significant association was observed under the dominant inheritance model (P =.039). Conclusion: CYP2R1 and CYP27B1 genes were over-expressed in CRC samples compared to the adjacent control tissues. Furthermore, CYP2R1 rs12794714 variant was associated with the risk of CRC in the studied samples. CYP2R1 rs10766196 and CYP27B1 rs4646536 are not responsible for CYP2R1 and CYP27B1 genes expression alteration, respectively, but CYP2R1 rs12794714 polymorphism may be the reason of CYP2R1 upregulation and increased the risk of CRC

The differential DNA hypermethylation patterns of 2 microRNA-137 and microRNA-342 locus in early 3 colorectal lesions and tumours

Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide, representing 13% of all cancers. The role of epigenetics in cancer diagnosis and prognosis is well established. MicroRNAs in particular influence numerous cancer associated processes including apoptosis, proliferation, differentiation, cell-cycle controls, migration/invasion and metabolism. MiRNAs-137 and 342 are exon- and intron-embedded, respectively, acting as tumour-suppressive microRNA via hypermethylation events. Levels of miRNAs 137 and 342 have been investigated here as potential prognostic markers for colorectal cancer patients. The methylation status of miRNA-137 and miRNA-342 was evaluated using methylation-specific (MSP) polymerase chain reaction (PCR) on freshly frozen tissue derived from 51 polyps, 8 tumours and 14 normal colon mucosa specimens. Methylation status of miRNA-137 and miRNA-342 was significantly higher in tumour lesions compared to normal adjacent mucosa. Surprisingly, the methylation frequency of miR-342 (76.3%) among colorectal cancer patients was significantly higher compared to miR-137 (18.6%). Furthermore, normal tissues, adjacent to the lesions (N-Cs), displayed no observable methylation for miRNA-137, whereas 27.2% of these N-Cs showed miRNA-342 hypermethylation. MiRNA-137 hypermethylation was significantly higher in male patients and miR-342 hypermethylation correlated with patient age. Methylation status of miRNA-137 and miRNA-342 has both diagnostic and prognostic value in CRC prediction and prevention