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.
Mammalian cell culture refers to culturing mammalian cells in a medium that provide nutrients for cells to be able to grow in vitro under environment that closely mimic the in vivo conditions. By enabling culturing these cells outside living biological entities, investigation on intra- and intercellular activities and flux; genetic and phenotyping analysis; proteomics, study of toxicology, drug discovery and development can be carried out without manipulation of living animals. In this chapter, detail protocol of media preparation, cell culture maintenance and preservation are elaborated for both types of mammalian cell culture, monolayer or suspension cultures. Determination of number of cells is discussed as well.
Aim: To determine the expression of tissue inhibitors of metalloproteinases (TIMP-2) in oral squamous cell carcinoma (OSCC) and the difference in its expression level between positive and negative HPV-16 (human papilloma virus- 16) OSCC patients. Methods: This study was conducted on 33 biopsies obtained from patients with OSCC and 10 normal oral mucosa as controls. In situ hybridization (ISH) was used to investigate the presence of HPV-16, while immunohistochemistry (IHC) was used to estimate the expression level of TIMP-2. Results: The TIMP-2 was expressed in 27 (81.8%) of OSCC sections with no significant difference between its expression level in HPV-16 positive and HPV-16 negative OSCC cases (p=0.058). TIMP-2 was found to be hig
... Show MoreBackground: Oral cancer is a highly lethal and disfiguring disease. Squamous cell carcinoma of the oral cavity constitutes about 90% of all oral malignancies. The aims of the study was to achieve an epidemiological description of the oral squamous cell carcinoma in Iraq in general and in Iraqi governorates except Kurdistan region retrospectively during period 2001-2013 Materials and Methods: Data were collected from department of oral and maxillofacial pathology, college of dentistry, university of Baghdad, Nuclear medicine and radiotherapy hospital, Iraqi cancer registry center, Main hospitals in Baghdad and Iraqi governorates, Private labs. for histopathological examinations. The descriptive and inferential statistical methods were used
... Show MoreBackground: Understanding the pathogenesis and molecular basis of Oral Squamous Cell Carcinoma (OSCC) has increased rapidly over the past few years that is essential to improve patient's prognosis and treatment modalities. The purpose of this study to evaluate the Immunohistochemical expressions of AKT, ATM, AND Cyclin E in oral squamous cell carcinoma Materials and methods: This study was performed on a forty formalin-fixed paraffin-embedded blocks which histopathologically diagnosed as Oral Squamous Cell Carcinoma. All cases were collected from the Histopathological Laboratory from patients treated surgically at Maxillofacial surgery Department at Ramadi Teaching Hospital, Iraq. Results: The immunohistochemical staining of AKT showed pos
... Show MoreBackground: Oral squamous cell carcinoma (OSCC) is a common malignancy characterized by poor prognosis and low survival rate. The purpose of this study was to evaluate the Immunohistochemical expressions of BAD, MDM2, and P21as apoptotic markers in oral squamous cell carcinoma. Materials and methods: This study was performed on forty formalin-fixed paraffin-embedded blocks which histopathologically diagnosed as Oral Squamous Cell Carcinoma. All cases were collected from the Histopathological Laboratory from patients treated surgically at Maxillofacial surgery Department at Ramadi Teaching Hospital, Iraq. Results: The immunohistochemical staining of BAD showed positive expression in 39 (97.5%), MDM2 showed positive expression in 39(97.5%)
... Show MoreBreast cancer is the second most common cancer in women world. Multiple Cytokines appear to have a dominant role in human breast cancer formation. Estimation of the in situ expression of IL-6 and IL-1β in breast cancer patients. A sixty patients with breast cancer BC were divided into two clinical subgroups, (30) with malignant breast cancer MBC and (30) with benign breast tumor as a control group according to histological examination. In situ hybridization technique used for detection of IL-6 and IL-1β mRNA sequence in two groups. The results showed that percentages of mRNA expression of IL-6 and IL-1β were in (≥ 11-50%) for malignant breast cancer. This research also investigated that (73.3%) of beni
... Show MoreGenerally, statistical methods are used in various fields of science, especially in the research field, in which Statistical analysis is carried out by adopting several techniques, according to the nature of the study and its objectives. One of these techniques is building statistical models, which is done through regression models. This technique is considered one of the most important statistical methods for studying the relationship between a dependent variable, also called (the response variable) and the other variables, called covariate variables. This research describes the estimation of the partial linear regression model, as well as the estimation of the “missing at random” values (MAR). Regarding the
... Show MorePrecise forecasting of pore pressures is crucial for efficiently planning and drilling oil and gas wells. It reduces expenses and saves time while preventing drilling complications. Since direct measurement of pore pressure in wellbores is costly and time-intensive, the ability to estimate it using empirical or machine learning models is beneficial. The present study aims to predict pore pressure using artificial neural network. The building and testing of artificial neural network are based on the data from five oil fields and several formations. The artificial neural network model is built using a measured dataset consisting of 77 data points of Pore pressure obtained from the modular formation dynamics tester. The input variables
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