Induction of Apoptosis in Human Cancer A549 Cells Through Hydroalcoholic Extract of Salvia officinalis
Volume 11, Issue 2, Summer 2020, Pages 100-112
https://doi.org/10.52547/JCT.11.2.100
E Hoveizi, F Pouratar, M Kesmati, A Shahriari
Abstract Aim: The purpose of the current study is to investigate the cytotoxic and oxidative effects of Salvia officinalis hydroalcoholic extract on cancer A549 cells.
Material and Methods: The cells were seeded in plates to investigate concentrations of Salvia officinalis extract and MTT measurement was done to determine IC50 concentration and cell viability on days 1, 3, 5, and 7. Moreover, analyses of superoxide dismutase and catalase enzymes involved in oxidative stress pathway, and AO/EB staining for a qualitative investigation of cell lines has been conducted in order to explore the effects of IC50 concentration on apoptosis induction. Also, Giemsa and DAPI stainings were utilized to explore the morphological changes of cell and nucleus..
Results: IC50 concentration of Salvia extract for A549 cells was determined 5 mg/mL. According to the results, Salvia extract reduced the cell viability of A549 cells. Based on the results, Salvia extract had a significantly greater cytotoxic effect compared to the control sample of A549 cell line. Treatment cells indicated some clear and dose-dependent differences at different times. The results of stainings proved the apoptosis induction in the treatment group. Furthermore, regarding the enzyme expression results the activity of superoxide dismutase and catalase enzymes in the cell groups treated by Salvia extract was significantly increased, compared to the control group.
Conclusion: Consistent with the results of this study, in addition to the anti-oxidative activity Salvia officinalis hydroalcoholic extract revealed significant apoptotic effects on lung cancer cells, considering a time and dose-dependent method.
Inhibition of lung cancer Calu-6 cell line proliferation using Cdc42 gene shRNA
Volume 8, Issue 3, Winter 2018, Pages 261-270
https://doi.org/10.52547/JCT.8.3.261
Z Ghambari, i M Nabiun, H Jalali, L Karimzadeh
Abstract Aim: In current study, we aimed to reduce Cdc42 gene expression in lung carcinoma related cells, Calu-6, and assayed its effect on cell proliferation.
Material and Methods: To reduce the expression of Cdc42 gene shRNA system was used and lentiviral system was selected to deliver Cdc42 specific shRNA to Calu-6 cells. Recombinant lentiviruses produced by co-transfection of pMD2G, psPAX2 and p-GFP-C-shLenti plasmids into 293T cells using lipofectamin. Efficiency of transfection and transduction assessed by florescent microscopy. Viability of cells treated by recombinant lentiviruses assessed by MTT assay.
Results: florescent microscopy showed 80% transfection of 293T cells and high rate of Calu-6 cells transduction. MTT assay results revealed that viability of transduced Calu-6 cells reached to %58 and %40 in compare to control and negative control cells, respectively.
Conclusion: recombinant lentiviruses properly transfer Cdc42-shRNA into Calu-6 cells, leading to reduction of cell proliferation. Silencing of Cdc42 gene expression using lentiviruses is persist and long-term effect which can be under attention for gene therapy of lung cancer.