جستجوی مقالات مرتبط با کلیدواژه "رده ی سلولی" در نشریات گروه "پزشکی"

In spite of the wide use, low proteome coverage and fuzzy patterns are the most important deterrents for the clinical applications of gel-based proteomic studies. So herein, we tried to increase the 2-dimentional proteome coverage of Hs578T breast cancer cells via investigating the efficacy of the three common techniques, usually used for interfering removal.
Hs578T cells were incubated in a lysis solution to obtain raw cell extracts. Cellular soups of each extraction were then pooled, homogenized, and aliquoted to be further treated by three different protein-specific purification methods including acetone, acetone-methanol, and trichloroacetic acid (TCA)-acetone, each in triplicates. All the purified protein pellets were then dissolved in a standard rehydration buffer solution, loaded into the 17-cm immobilized pH gradient (IPG) strips, and separated according to their isoelectric points. Proteins were then separated once more according to their molecular weights in an OFarrell separation system. Finally, by the visualization of the protein spots on the 2-dimentional profiles, quality and quantity of these 2-dimentional proteome patterns were then analyzed using the ImageMaster software.
Findings: The obtained proteome recovery yields and total protein counts for acetone, acetone-methanol, and trichloroacetic acid-acetone methods were 0.100 ± 0.001, 0.070 ± 0.002, and 0.120 ± 0.005 ng/cell, and 1299 ± 9, 1698 ± 14 and 1973 ± 17, respectively. The results represent data obtained from three independent experiments.
Trichloroacetic acid-acetone purification not only represented the highest recovery yield, suitable for expensive assays, but also showed the most suitable proteome coverage. So, the method is recommended for the comparative proteomic studies. However, the acetone-methanol procedure is more recommended for serological proteome analysis (SERPA); since it represents stronger protein spots which are more fitted to the immunoblotting procedure.

Cancer-Testis genes (CT-genes) are a gene family that only expressed in normal testis tissue and some of them are randomly expressed in some types of cancers. These genes can be promising cases for immunotherapy of breast cancer. This research was carried out for comparison of the expression frequencies these genes in cancerous cell lines and tumor samples. In this analytical-descriptive study, after providing three breast-cancer cell lines and their cultures in appropriate medium, RNA extraction and cDNA synthesis were done and expressions of NY-ESO-1 1a, NY-ESO-1 1b, SCP1, SSX-2 and MAGE-3 genes were studied using multiplex real time-polymerase chain reaction (multiplex RT-PCR) method. None of cell lines expressed Cancer-Testis genes; but all of them expressed glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene (internal control). According to the role of these genes in gametogenesis, we can consider that their expressions in cancer cells are based on dedifferentiation. The observed difference between expression frequency of these genes in breast-cancer cell lines and tumor samples can be related to several passages which are carried out for providing cell lines. By examining more cell lines of breast cancer for these genes, we can achieve more accurate results.

Mycoplasma is a major contaminant of cell lines and is cosiderd as a serious problem of economic and biological importance in basic research, diagnosis, and biotechnology products. Detection of mycoplasma infection in cell cultures started on microbiological culture; later, other methods like DAPI staining and serological tests such as Indirect Immunofluorescnse, ELISA, DNA probe, PCR, PCR-ELISA, and Real-Timr PCR developed for detection of mycoplasma. In this study, a sensitive, specific, and rapid method was used for detection of varity of mycoplasma species in cell lines. This method was based on a PCR reaction using genus specific primers for 11 mycoplasma species. Mycoplasma contamination using this assay was examind for 183 different cell line deposited in national cell bank of Iran. PCR showed that 48.6% of cell lines were contaminated with mycoplasma while 27.3% of them were found to be infected. In comparison to microbiological culture, PCR method was shown to be 100% sensitive and 70.7% specific. Our results using species specific primers reveald that the most important contaminating mycoplasma species in cell lines were mycoplasma fermentans, mycoplasma arginini, mycoplasma hyorhinis, and mycoplasma orale. We were also able to identify a number of cell lines which were contaminated whit more than one species of mycoplasma.