Quantitative real-time Polymerase Chain Reaction (RT-qPCR) has become a valuable molecular technique in biomedical research. The selection of suitable endogenous reference genes is necessary for normalization of target gene expression in RT-qPCR experiments. The aim of this study was to determine the suitability of each 18S rRNA and ACTB as internal control genes for normalization of RT-qPCR data in some human cell lines transfected with small interfering RNA (siRNA). Four cancer cell lines including MCF-7, T47D, MDA-MB-231 and Hela cells along with HEK293 representing an embryonic cell line were depleted of E2F6 using siRNA specific for E2F6 compared to negative control cells, which were transfected with siRNA not specific for any gene. Using RT-qPCR, Ct (threshold cycle) values of 18S and ACTB were determined in transfected cells and compared with control cells. In the selection of the above cell lines, 18S was identified as the most stably expressed reference gene than ACTB in gene knockdown experiments.
Background: Previous studies about the correlation of genetic polymorphisms in the multigene family of cyto- chrome P450 (CYPs), the effect of tobacco smoking, and the risk of developing cancer have been well in- vestigated in different populations, but not in Iraq. Furthermore, the studies of malignance occurrence re- lationship with cigarette tobacco smoking revealed the presence of strong association, however, little is known about the risk of Waterpipe (WP) tobacco smoking. Thus, determination two important genetic polymorphisms in CYP1A1, a main member of CYPs, among Iraqi men was our first aim. This is the first study that highlights the correlation of CYP1A1 polymorphisms with the risk of lung cancer in Iraq. The second aim was to ev
... Show MoreBackground: Metabolic syndrome (Mets) is partially heritable. High mobility group AT-hook1 (HMGA1), an architectural transcription factor, affects the homeostasis of glucose. The marked inter-individual differences between T
... Show MoreCeliac disease (CD) is an autoimmune disorder characterized by chronic inflammation that essentially affects the small intestine and is caused by eating gluten-containing foods. This study sought to determine gene expression of NLRP3 Inflammasome in peripheral blood of Iraqi CD children using quantitative real-time PCR (qRT-PCR) assay. Thirty children with CD (12 males and 18 females) were enrolled in the study and their age range was 3-15 years. The diagnosis of the disease was confirmed by serological examinations and intestinal endoscopy. A control sample of 20 age-matched healthy children was also included. The children were stratified for age, gender, body max index (BMI), histological findings, and marsh classification. Furthe
... Show MoreAbstract: E2F6 is a member of the E2F family of transcription factors involved in regulation of a wide variety of genes through both activation and repression. E2F6 has been reported as overexpressed in breast cancers but whether or not this is important for tumor development is unclear. We first checked E2F6 expression in tumor cDNAs and the protein level in a range of breast cancer cell lines. RNA interference-mediated depletion was then used to assess the importance of E2F6 expression in cell lines with regard to cell cycle profile using fluorescence-activated cell sorting and a cell survival assay using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The overexpression of E2F6 was confirmed in breast tumor cDNA samp
... Show MoreThe current study was conducted in the period extending from November 2018 to October 2019 and designed as a case-control study and aimed to assess the seroprevalence of HCMV. However, a total number of 91serum specimens were collected to fulfill this purpose from females (71 breast cancer patients, and control group of 20 females) attending Al-Amal hospital for cancer management and Baghdad teaching hospital and the practical part was performed in College of Science, University of Baghdad. The study protocol was approved by the Ethics Committee at the Department of Biology (Reference: BEC/0220/0011). The immunological part for evaluation of seroprevalence of HCMV was accomplished by ELISA technique which revealed that anti-HCMV IgG was sco
... Show MoreBackground: Chronic myelogenous leukemia is a malignant hematological disease of hematopoietic stem cells. It is difficult to adapt treatment to each patient's risk level because there are currently few clinical tests and no molecular diagnostics that may predict a patient's clock for the advancement of CML at the time of chronic phase diagnosis. Biomarkers that can differentiate people based on the outcome at diagnosis are needed for blast crisis prevention and response improvement. Objective: This study is an effort to exploit the SLC25A3 gene as a potential biomarker for CML. Methods: RT-qPCR was applied to assess the expression levels of the SLC25A3 gene. Results: In comparison to the mean ΔCt of the control group, which was found to b
... Show MoremiRNAs regulate protein abundance and control diverse aspects of cellular processes and biological functions in metabolic diseases, such as obesity and diabetes. Lethal-7(Let-7) miRNAs specifically target genes associated with diabetes and have a role in the regulation of peripheral glucose metabolism. The present study aimed to describe the gene expressions of the let-7a gene with the development of diabetes in Iraq and the difference in the expression of this gene in patients with diabetes and healthy individuals. The association between age and gender with the development of diabetes was studied in this study and the results were compared with those of healthy individuals in the group of control. Based on the obtained results, there was
... Show MoreThis study dealt with IL-13 1024 (C/T) gene genotyping among patients with Thyroid goiter in Iraq. Forty blood samples from patients with Thyroid goiter were collected and compared with 30 healthy persons as controls. The genotyping results of IL-13 1024 (C/T) gene using ARMS-PCR revealed presence TT, CC and CT genotypes beside T and C alleles. The T allele and TT genotype frequency were higher in Thyroid goiter patients compared to the same genotype and allele in healthy persons (P = 0.060). These increasing results were related with increasing risk factor of Thyroid goiter (odds ratio [OR] 2.15; 95% confidence interval [CI] 0.99–71.4). No significant differences between genotypes for Thyroid goiter patients and controls were revealed by
... Show MoreThe effect of the aqueous extract of fenugreek seeds (Trigonella Foenum Graecum L.), Rhodium complex (?) with formula [RhL2CLH2O].1 1/2 ETOH and palladium (?) [pdl2].2ETOH,where L=2-hydroxy phenyl piperonalidine was studied on two cancer cell lines. The first cell line was intestine cancer of female albino mice (L20B), the second one was Rhabdomysarcomas (RD)cell line in human. The activity of the new complexes and the aqueous extract was compared to the well-known anticancer drug (cis-platin) by utilizing the in vitro system. The cell lines were treated with four concentrations of cis-platin 31.25,62.5,125 and 250 ?g/ml for 72 hour exposure time. The same concentrations were used with extract and the new complexes. This study showed that t
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