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Showing 3 results for Cell Survival
In vitro selective growth inhibition of breast adenocarcinoma cell lines by Pseudomonas sp. UW4 metabolites
Maedeh Pasiar , Leila Rouhi , Zahra Bamzadeh , Seyed Hossein Hejazi ,
Volume 74, Issue 9 (12-2016)
Abstract

Background: Breast cancer is a malignant proliferation of epithelial cells that lining the ducts or lobules of the breast. It is the second common cancer, after lung cancer in women. Since growth inhibition is an important strategy in cancer treatment, many attempts are in program to find new apoptotic inducer agents. Today there is some reports about effect of metabolites of Pseudomonas on cancer cells, hence, metabolites of Pseudomonas sp. UW4, were isolated and anti-cancer and anti-microbial activity of these metabolites was studied.

Methods: This experimental study was performed in cellular and developmental biology of Shahrekord Islamic Azad University from April 2015 to August 2015. Anti-microbial activity of metabolites of Pseudomonas sp. UW4 was tested against a pathogenic bacteria, including Escherichia coli, Bacillus cereus and Staphylococcus aureus. For anti-cancer activity, in this study SKBR3 cells and normal fibroblast cells (HU-02) were cultured in DMEM medium with 10% fetal bovine serum (FBS). The cells were treated by various concentrations of these metabolites 5, 10, 15 and 20 mg/ml for 24, 48 and 72 h. Cell viability was assessed by MTS assay. Cells were seeded at 5×103 cells/ml in 96 well plates and incubated for 24 hr. Then metabolites of bacteria were added, after indicated times MTS (20 µl) was added and the absorbance was measured at 492 nm using ELISA plate reader.

Results: Pseudomonas sp. UW4 was able to produce antimicrobial metabolites against Staphylococcus aureus. Metabolites decreases the viability of SKBR3 cell line in a time and dose dependent manner, so that the most effective concentration of this substance was 20 mg/ml and 72 h after treatment (P< 0.01). While Pseudomonas sp. UW4 in various concentrations had no significant effect on normal fibroblast cells (P= 0.24).

Conclusion: Bioactive compounds produced by of Pseudomonas sp. UW4 could be used for elimination of infections and treatment of breast cancer SK-BR3.

Effect of miRNA-1266-5p repression on the increasing cell survival and alterations of the cell cycle in AGS cell line
Saba Sorayyayi , Sogand Vahidi , Mohammad Mohammadzadeh , Sayyed Saied Hosseini-Asl ,
Volume 76, Issue 8 (11-2018)
Abstract
Background: Gastric cancer is among the most common malignancies in certain parts of the world, such as northwest Iran. miRNAs are small and single-stranded noncoding RNAs with about 19-23 nucleotides. Several studies have shown that miRNAs play important roles in gastric tumorigenesis. The aim of this study was to determine the effect of miRNA-1266-5p repression on the cell survival and alterations of the cell cycle in gastric cancer cell line of AGS (NCBI Code: C131, Gastric epithelial cell line).
Methods: This experimental study was performed from April to December 2017 in Cellular-Molecular Research Center of Ardabil University of Medical Sciences, Iran. In this study, AGS cells were cultured in RPMI-1640 medium containing 10% serum and 1% antibiotic. The cells were transfected with miR-1266-5p mimic, miR-1266-5p inhibitor and HiPerFect reagent alone as negative control. The miR-1266-5p expression and transfection efficiency were analyzed by Stem-loop TaqMan qRT-PCR. The cell proliferation and cell cycle alterations were determined using MTT calorimetric assay and flow cytometry, respectively. The results were analyzed using SPSS 19.0 statistics software (SPSS Inc., Chicago, IL, USA) and presented as the means±standard deviation (SD).
Results: miR-1266-5p expression was increased in AGS cells transfected with miR-1266-5p mimic compared to control cells (P=0), while miR-1266-5p expression was decreased in transfected cells with the inhibitor compared to controls (P=0). Among different time points, the most effects of miR-1266-5p mimic and inhibitor were noticed after 48 hours of transfection. The upregulated miR-1266-5p significantly decreased cell growth, in contrast, inhibitor promoted cell proliferation (P=0). In addition, miR-1266-5p upregulation induced cell cycle arrest at the transition of G1 to S phase and led to G0/G1 entry (P=0), while of miR-1266-5p led to G2/M entry (P=0.001).
Conclusion: According to the results obtained from this study, miR-1266-5p can reduce cell survival and induce cell cycle arrest and act as a tumor suppressor in AGS cells. While its inhibition can increase cell survival and reduce apoptosis.

Comparison of MTT, trypan blue, and clonogenic assay, to determine the viability in human anaplastic thyroid cancer cell line
Marjan Ghorbani-Anarkooli , Sara Dabirian, Hasan Moladoust, Adib Zendedel, Mohammad Hadi Bahadori,
Volume 77, Issue 1 (4-2019)
Abstract
Background: Evaluation of cell viability is momentous in pharmacologic and oncological research. Cell viability evaluation determines cell sensitivity and consequently treatment outcome. Various methods are available to determine cell survival. Each of these methods evaluates different endpoints. Accordingly, determining the correlation between these methods is important. In this study, in order to determine the viability of human anaplastic thyroid cancer cell line, the sensitivity of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, trypan blue test and clonogenic assay were compared.
Methods: This experimental study was performed in the Cellular and Molecular Research Center at Guilan University of Medical Sciences, Rasht, Iran from October 2016 to March 2017. The human anaplastic thyroid cancer cell line was cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS). The cultured cells were treated with melatonin, for 24 hours. Then, the viability of the cells was evaluated by MTT assay, trypan blue test and clonogenic assay. Furthermore, plating efficiency and surviving fraction were used in order to draw survival curve in the clonogenic assay.
Results: The concentration of melatonin at IC50 point was 4.794±0.117 millimolar (mM) in MTT assay, 4.375±0.894 mM in trypan blue test and 2.246±0.326 mM in clonogenic assay. Comparing the IC50 values of these test revealed that C50 values obtained from MTT assay and trypan blue test had no significant difference (P=0.6446), while there was a significant difference between IC50 values obtained from MTT and clonogenic assays (P=0.0032). Moreover, the IC50 values obtained from trypan blue test and clonogenic assay were also significantly different (P=0.0078). The results of the regression analysis of cell viability were shown a linear, positive and significant correlation between these three methods and MTT assay and trypan blue test showed higher correlation (r=0.99, P<0.001).
Conclusion: Based on our results, all these methods were effective to identify cytotoxicity in human anaplastic thyroid cancer cell line, while MTT assay and trypan blue test were more sensitive than clonogenic assay.

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