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.
Angiogenesis is important for tissue during normal physiological processes as well as in a number of diseases, including cancer. Drug resistance is one of the largest difficulties to antiangiogenesis therapy. Due to their lower cytotoxicity and stronger pharmacological advantage, phytochemical anticancer medications have a number of advantages over chemical chemotherapeutic drugs. In the current study, the effectiveness of AuNPs, AuNPs-GAL, and free galangin as an antiangiogenesis agent was evaluated. Different physicochemical and molecular approaches have been used including the characterization, cytotoxicity, scratch wound healing assay, and gene expression of VEGF and ERKI in MCF-7 and MDA-MB-231 human breast cancer cell line. Re
... Show MoreThere is an assumption implicit but fundamental theory behind the decline by the time series used in the estimate, namely that the time series has a sleep feature Stationary or the language of Engle Gernger chains are integrated level zero, which indicated by I (0). It is well known, for example, tables of t-statistic is designed primarily to deal with the results of the regression that uses static strings. This assumption has been previously treated as an axiom the mid-seventies, where researchers are conducting studies of applied without taking into account the properties of time series used prior to the assessment, was to accept the results of these tests Bmanueh and delivery capabilities based on the applicability of the theo
... Show MoreMammalian 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.
In the present study, five derivatives have been designed to be synthesized as possible mutual prodrugs for 5-Fluorouracil (5-FU) and non steroidal anti-inflammatory drugs (NSAIDs) to selectively deliver the drugs into the cancer cells. The synthesis of the target compounds were accomplished following multistep reaction procedures, the chemical reaction followed up and the purity of the products were checked by TLC. The structure of the final compounds and their intermediates were confirmed by their melting points, infrared spectroscopy and elemental microanalysis, the hydrolysis of compound III was studied using HPLC technique. According to the results mentioned above, compounds (I−V) can be good candidates as possible mutual prod
... 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 MoreSince the beginning of the last century, the competition for water resources has intensified dramatically, especially between countries that have no agreements in place for water resources that they share. Such is the situation with the Euphrates River which flows through three countries (Turkey, Syria, and Iraq) and represents the main water resource for these countries. Therefore, the comprehensive hydrologic investigation needed to derive optimal operations requires reliable forecasts. This study aims to analysis and create a forecasting model for data generation from Turkey perspective by using the recorded inflow data of Ataturk reservoir for the period (Oct. 1961 - Sep. 2009). Based on 49 years of real inflow data
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