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.
This study involved the effect of the aqueous extracts of two plants, Origanum vulgare L.(1), Trigonella Foenum Graecum L. (Fenugreek) seeds(2) on the growth of cancer cell lines. Rhabdomyo sarcomas (RD) of human cell line and female intestine cells of Albino mice (L20B) in vitro System. These extracts were compared with the known anticancer drug Cis-platinum(Cis-Pt) as a positive control. The phytochemical tests were used for screening the active compounds in plants. The inhibition activity assay was used as a parameter of the cytotoxic effect of these extracts. Cancer 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 for the other ext
... Show MoreThis study was aimed to determine the mutations and single nucleotide polymorphisms (SNPs) in exon 3 and 7 of estrogen receptor beta (ESR2) gene in women with breast cancer from Iraq. Different samples (blood, fresh tissue with blood from same patient, and formalin fixed paraffin embedded, FFPE) were collected from women with breast cancer. Molecular analysis exon 3 and 7 in ESR2 has been studied by using PCR. It was found exon 3 and 7 in ESR2 were revealed as a single band with size 151 and 157 bp, respectively. There was no SNP in exon 3 has been identified. While three novel polymorphisms (ACT, AGG and GCA) were detected in exon 7, the type of those polymorphisms deletion for ACT and AGG while substitution polymorphism for GCA. From this
... Show MoreThe cytotoxic effect of catechol was examined in two human cancer cell lines, Epidermoid larynx carcinoma (Hep- 2), Cerebral glioblastoma multiforme (AMGM-5) and Murine mammary adenocarcinomacell (AMN3) treated with half concentrations of catechol (1000, 500, 250, 125, 62.5 and 32.25 μM) for 72 hr. The get hold of results showed catechol have a toxic effect of the cell viability of three types of cell lines after 72h of exposure, the toxicity was dependent on catechol concentrations and/or autoxidation for quinines formation, there were a marked decreased of cell viability in a dose dependent manner in all cell line types. Inhibition concentration of catechol for 50% of cell viability (IC50) were calculated, they were at 581.5 μM, 478 μM
... Show Moreفي هذا البحث تم تحضير المركبات المعدنية الجديدة لأيونات البلاتين (الرباعي) و الذهب (الثلاثي) مع ليكاند قاعدة مانخ جديد مشتق من السيبروفلوكساسين . تم استخدام المعقدات بعد ذلك كمصدر لتحضير جزيئات عن طريق ترسيب المعقدات على مسام دقائق السيليكا النانوية. Si/Au2O3 Si/PtO2 تم تشخيص الليكاند و معقداته
... Show MoreGenetic polymorphisms of genes whose products are responsible for activities, such as xenobiotic metabolism, mutagen detoxification and DNA-repair, have been predicted to be associated with the risk of developing lung cancer (LC). The association of LC with tobacco smoking has been extensively investigated, but no studies have focused on the Arab ethnic- ity. Previously, we examined the association between genetic polymorphisms among Phase I and Phase II metabolism genes and the risk of LC. Here, we extend the data by examining the correlation of OGG1 Ser326Cys combined with CYP1A1 (Ile462Val and MspI) and GSTP1 (Ile105Val and Ala103Val) polymorphisms with the risk of LC. Polymerase chain reaction- restriction fragment length polymorphism (
... Show MoreIn this research, silver nanoparticles (AgNPs) were manufactured using aqueous extract of mushroom Pleurotus ostreatus. Anticancer potential of AgNPs was investigated versus human breast cancer cell line (MCF-7). Cytotoxic response was assessed by MTT assay. AgNPs showed inhibition effect at the following concentrations 12.5, 25, 50, 100 and 200 µg/ml versus MCF-7 cell line, and all treatments had a positive result. The MCF-7 cells were inhibited up to 85.14 % at the concentration 200 μg/ml of AgNPs which reduced cells viability to 14.86%, while 12.5 μg/ml of AgNPs caused 24.23% cells inhibition with reduction of cells viability to 75.77%.
Coagulation is the most important process in drinking water treatment. Alum coagulant increases the aluminum residuals, which have been linked in many studies to Alzheimer's disease. Therefore, it is very important to use it with the very optimal dose. In this paper, four sets of experiments were done to determine the relationship between raw water characteristics: turbidity, pH, alkalinity, temperature, and optimum doses of alum [ .14 O] to form a mathematical equation that could replace the need for jar test experiments. The experiments were performed under different conditions and under different seasonal circumstances. The optimal dose in every set was determined, and used to build a gene expression model (GEP). The models were co
... Show MoreDespite extensive investigations, an effective treatment for sepsis remains elusive and a better understanding of the inflammatory response to infection is required to identify potential new targets for therapy. In this study we have used RNAi technology to show, for the first time, that the inducible lysophosphatidylcholine acyltransferase 2 (LPCAT2) plays a key role in macrophage inflammatory gene expression in response to stimulation with bacterial ligands. Using siRNA- or shRNA-mediated knockdown, we demonstrate that, in contrast to the constitutive LPCAT1, LPCAT2 is required for macrophage cytokine gene expression and release in response to TLR4 and TLR2 ligand stimulation but not for TLR-independent stimuli. In addition, cells transfe
... Show MoreOut of 120 isolates from different clinical cases, only 75 were found and confirmed that they belong to the Pseudomonas aeruginosa bacteria. The result revealed that the LasB virulent gene was present in 63 isolates with 63% percentage. The gel electrophoresis showed that the molecular weight of LasB gene was 300 bp. DNA sequences of LasB gene was done, and the results showed the presence of some gene mutations like substitution, addition and deletion with 97% identity with the Refseq gene. From the other side, the results of identities of translated nucleotides sequence with the original sequence of amino acids revealed that there are no effects of gene mutations on translation of the product protein.
Epithelial and stromal communications are essential for normal uterine functions and their dysregulation contributes to the pathogenesis of many diseases including infertility, endometriosis, and cancer. Although many studies have highlighted the advantages of culturing cells in 3D compared to the conventional 2D culture system, one of the major limitations of these systems is the lack of incorporation of cells from non‐epithelial lineages. In an effort to develop a culture system incorporating both stromal and epithelial cells, 3D endometrial cancer spheroids are developed by co‐culturing endometrial stromal cells with cancerous epithelial cells. The spheroids developed by this method are phenot