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.
Leishmania species are the causative agent of a tropical disease known as leishmaniasis. Previous studies on the old world species Leishmania major, showed that the amastigotes form which resides inside the macrophage of the vertebrate host, utilize host’s sphingolipids for survival and proliferation. In this study, gene expression of serine palmitoyltransferase (SPT) subunit two (MmLCB2) of the mouse macrophage cell line (RAW264.7), which is the first enzyme in the de novo sphingolipid biosynthesis, was detected in both infected and non-infected macrophages. This was detected under condition where available sphingolipid was reduced, with the new world species Leishmania mexicana. Results of qPCR analysis showed that there was no differen
... Show MoreBackground: CYP1A1 gene polymorphisms and tobacco smoking are among several risk factors for various types of cancers, but their influence on breast cancer remains controversial. We analyzed the possible association of CYP1A1 gene polymorphisms and tobacco smoking-related breast cancer in women from Iraq. Materials and methods: In this case-control study, gene polymorphism of CYP1A1 gene (CYP1A1m1, T6235C and CYP1A1m2, A4889G) of 199 histologically verified breast cancer patients' and 160 cancer-free control women's specimens were performed by using PCR-based restriction fragment length polymorphism. Results: Three genotype frequencies (TT, TC, and CC) of CYP1A1m1T/C appeared in 16.1, 29.6, and 54.3% of women with breast cancer, respectiv
... Show MoreThis study focused on the expression and regulation of BRCA1 in breast cancer cell lines compared to normal breast. BRCA1 transcript levels were assessed by real time quantitative polymerase chain reaction (RT-qPCR) in the cancer cell lines. Our data show overexpression of BRCA1 mRNA level in all the studied breast cancer cell lines: MCF-7, T47D, MDA-MB-231 and MDA-MB-468 along with Jurkat, leukemia T-lymphocyte, the positive control, relative to normal breast tissue. To investigate whether a positive or negative correlation exists between BRCA1 and the transcription factor E2F6, three different si-RNA specific for E2F6 were used to transfect the normal and cancerous breast cell lines. Interestingly, strong negative relationship was found b
... Show MoreThe present study conducted on 120 obese males and 50 healthy males, their age ranged from 20-50 years. The patients were divided into 3 groups based on Body Mass Index (BMI) and Central Obesity (CO), it has noticed that there is a significant relation between both indexes. The DNA was isolated from the blood of patients and applies for PCR by using designed primers for exons 1 and 3 of GCG gene. The results showed that there are mutants in exon 1 at locus 9573 (G/C) for 30 patients and locus 9864 (C/-) for 10 patients with X2 = 12.30, also it has fund mutants in exon 3 at locus 5397 (A/G) for 28 patients and locus 5434 (G/A) for 8 patients with X2 = 11.11. These mutants have a high significant effect P≤ 0.001 to cause pathogenicity.
... Show MoreThis study was conducted to use the local Ephedra alata plant as a model for extracting and detecting alkaloids in the stem of plant (alkaloids-rich extract and crude extract). Different extraction procedures were adopted for qualitative as well as the quantitative examination of the alkaloid extracts, as well as plant crude extract, the best methods for the extraction of the plant materials were applied. Simple, fast and accurate methods like TLC (thin layer chromatography) and HPLC (High-performance liquid chromatography), were used for the identification of the alkaloids (ephedrine) in different extracts of stems E. alata stems. Ephedrine alkaloid was detected in each alkaloids-rich and crude extrac
... Show MoreA field experiment was carried out in the College of Agricultural Engineering Sciences - University of Baghdad, during the fall season of 2021 to find out which cultivated cultivars of maize are efficient under nitrogen fertilization. The experiment was applied according to an RCBD (split-plot design with three replications). The cultivars of the experiment (Baghdad, 5018, Sarah) supply three levels of nitrogen fertilizer, which are N1 (100 kg.N/ha), N2 (200 kg.N/ha) and N3 (300 kg.N/ha). The statistical analysis results showed the superiority of the Sarah genotype, which gave the highest value of SOD and CAT enzymes, reaching 11.59 units mg-1 and 10.76 units mg-1 . Protein sequentially, while cultivar5018 outperformed as it gave th
... Show MoreCurrently, there is a growing interest in medicinal plants extracts as some plants have shown antitumor potential. The goal of this study was to test the anticancer activity of methanol extract of Conocarpous erectus leaves in breast cancer cells. Cytotoxicity was tested in vitro on breast cancer cell lines, MCF7 [Estrogen receptor + (ER+)] and MDA-MB231 [Estrogen receptor - (ER-)], in addition to normal fibroblast cells (REF). MTT assay was utilized to measure the growth inhibitory effects after 48 hours exposure to extracts. Viability results indicated that MDA-MB231 were sensitive (GI50 = 56.1µg/ml).However, no sensitivity was seen in both MCF7 and REF cells (GI50>100 µg/ml). I
... Show MoreGlutathione-S-transferases (GSTs) play a role in the detoxification of environmental chemicals and mutagens, such as those inhaled during tobacco smoking. There have been conflicting reports concerning GST polymorphisms as risk factors in the development of lung cancer. No studies focused on Arab populations exposed to Waterpipe (WP) tobacco smoke have been undertaken. Here Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and gene sequenc- ing were applied to analyze allelic variations in GSTP1-rs1695 and -rs1138272 amongst 123 lung cancer patients and 129 controls. The data suggest that WP smoking raised the risk of lung cancer more than three-fold (OR 3.6; 95% CI 2.1–6.0; p < 0.0001). However, there was no s
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