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: Carcinoma of urinary bladder is one of the most common malignancies worldwide and constitutes a major health problem. Multiple risk factors are associated with this tumor and its prognosis will depend on different clinicopathological parameters. Over expression of P53 protein and mutant Rb gene is associated with more aggressive clinical and histopathological features of the tumor such as advanced stage and higher grade. AIM: The immunohistochemical expression of Rb gene and P53 gene will be assessed through their protein products in transitional cell carcinoma (TCC) of the urinary bladder and then will be correlated with other well-known risk factors and prognostic parameters of bladder TCC, such as grading, tumor size, smoking
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The aim of this study was to establish the existence and interaction of TMPRSS2 – ERG gene fusion status with clinicopathological features of prostate cancer patients. This research consisted of 123 embedded formalin-fixed tissues obtained from the prostate tumor patients. The above gene fusion is detected through the technique of fluorescent in situ hybridization (FISH) by means of a triple color probe. Seven samples have not been scored due to technical difficulties and 46 patients have fusion (39.6%), while the remaining (70) have not been seen with fusion. Of the 46 fusion-positive, 17 (36%) were caused by ERG-translocation, of the other 29 (63%) were caused by the interstitial segment deletion between the two genes due to the
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Background: Peripheral giant cell lesion (PGCL) and central giant cell lesion (CGCL) of the jaws have a distinct clinical behavior.Giant cell tumour (GCT) is a benign locally aggressive neoplasm affects the long bones. Both lesions are characterized histologically by multinucleated giant cells in a background of ovoid to spindle-shaped mesenchymal cells. The WW domain-containing oxidoreductase (WWOX) gene is located at 16q23.1–16q23.2, a region that spans the second most common human fragile site, FRA16D, at 16q23.2.The Ki-67 antigen is a nuclear protein that is associated with and may be necessary for cellular proliferation.Ki-67 protein is present during all active phases of the cell cycle (G1, S, G2, and mitosis), but is absent fr
... Show MoreAnchusa strigosa - prickly alkanet from Boraginaceae grows in roadsides, and fields of a broad range of habitats from mediterranean woodlands, to steppe vegetation, to true desert. It is commonly known as" him him" or "lisan al thawr". Anchusa can withstand hard weather conditions and hence is widely cultivated. The color of its flowers can range from pure white to deep cobalt blue. Various parts of A. strigosa are used in traditional medicine for treating several diseases or symptoms, such as abdominal pain, bronchitis, cough, and diarrhea. The goal of this study was to examine the cytotoxic effect of the crude extract of A. strigosa roots and leaves and their fractions against various tumor cell lines: adenoc
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FLI1 is a member of ETS family of transcription factors that regulate a variety of normal biologic activities including cell proliferation, differentiation, and apoptosis. The expression of FLI1 and its correlation with well-known breast cancer prognostic markers (ER, PR and HER2) was determined in primary breast tumors as well as four breast cancer lines including: MCF-7, T47D, MDA-MB-231 and MDA-MB-468 using RT-qPCR with either 18S rRNA or ACTB (β-actin) for normalization of data. FLI1 mRNA level was decreased in the breast cancer cell lines under study compared to the normal breast tissue; however, Jurkat cells, which were used as a positive control, showed overexpression compared to the normal breast. Regarding primary breast carcinoma
... Show MoreProduction and characterization of methionine γ- lyase from Pseudomonas putida and its effect on cancer cell lines
In latest decades, genetic methods have developed into a potent tool in a number of life-attaching applications. In research looking at demographic genetic diversity, QTL detection, marker-assisted selection, and food traceability, DNA-based technologies like PCR are being employed more and more. These approaches call for extraction procedures that provide efficient nucleic acid extraction and the elimination of PCR inhibitors. The first and most important stage in molecular biology is the extraction of DNA from cells. For a molecular scientist, the high quality and integrity of the isolated DNA as well as the extraction method's ease of use and affordability are crucial factors. The present study was designed to establish a simple, fast
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