Cell and Tissue Journal

Abstract Aims: Nanoparticles due to their wide applications in medicine,industry,and biotechnology, have attracted many scientists’ attentions. Recently, nanoparticles especially selenium nanoparticles are widely used to diagnosis and cancer treatment. The aim of this study was to evaluate the cytotoxic and anticancer effects of selenium nanoparticles on colon cancer cell line and analysis of CAD (Caspase Activated DNase) gene expression.
Material and methods: In this study, colon cancer HT29 and normal HEK293 cell lines were purchased from the Pasteur Institute Cell Bank of Tehran and treated with selenium nanoparticles overnight. The cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Scotland) medium with 10% FBS serum and 1% streptomycin antibiotic (Gibco, Scotland). The cells were then stored at 37 ° C. In this study, cytotoxic effect of Selenium NPs was evaluated on HT29 and HEK293 cells using MTT (3-(4, 5-Dimethyltetrazollium Bromide) assay. Subsequently, they were treated with selenium nanoparticles in different concentrations (0, 7.81, 15.62, 31.25, 62.5, 125, 250 and 500 mg/mL) for 24 hours. To solubilize the viable cells formazan crystals production, we added 100 μl/well of dimethyl sulfoxide (DMSO) to them. After treatment of HT29 cells with IC₅₀ concentration, the total RNA was extracted and cDNA synthesized. Moreover, CAD gene expression was evaluated using Real Time PCR method. The data was evaluated by ABI StepOne utilizing the Applied Biosystems qRT-PCR (ABI 7300 system, Applied Biosystems). The quantification of the mode of Selenium NPs -induced cell death in the HT29 cells were ascertained using flow cytometry followed by staining with fluorescein isothiocyanate (FITC)‐Annexin V and propidium iodide (PI) staining. Finally, the study of apoptosis and necrosis of Selenium NPs was evaluated using flow cytometry method. Data analysis was statistically determined by using One-way analysis of variance (ANOVA) with SPSS/22 software followed by a Tukey test.
Results: The result showed that the treatment of Selenium NPs at 31.25 to 500 µg/mL concentration had maximum cytotoxic effect, revealed statistically significant (P˂0.001). The IC₅₀ value for Selenium NPs were measured at 75 µg/mL after 24 hours. In order to determine the effect of Selenium NPs on cancerous cells, alterations in the mRNA expression levels of CAD gene in HT29 cells were done by qRT-PCR technique followed by the exposure to nanoparticle. The CAD gene expression comparing to reference gene was up-regulated 4.04±0.125 fold. To determine the mechanism of cell death in the cancer cells, annexin V/PI flow cytometry was carried out. In the treatment of HT29 cells by IC₅₀ of selenium NPs, 10.43%, and, 24.28% of early and late stages’ apoptosis were observed, respectively
Conclusion: Our results suggest that selenium NPs can display some promising cytotoxic properties through inducing apoptosis pathway. Based on the results, up-regulated gene expression involved in apoptosis (CAD) and activating apoptosis, it can be concluded that the selenium NPs can be used as drug candidate in colon cancer treatment, but more studies are needed regarding the medicinal importance of nanoparticles.

Abstract Aim: The aim of the current work was to investigate the anticancer activities of Zinc Oxide nanoparticles (ZnONPs) through modulation of Glutation peroxidase and Glutationreductase gene expression in breast cancer cells.
Material and Methods: In this investigation, the breast cancer MCF-7 and normal HEK293 cell lines were treated with various concentrations of ZnONPs for overnight. Cell viability was measured using MTT(3-(4, 5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay against cancer and normalcells. The quantity of Glutation peroxidase and Glutationreductase genes compared to GAPDH gene expressions were evaluated using the real time PCR method.
Results:The MTT data showed that ZnONPs significantly decreased the viability of cancer cells compared to normal cells in dose-dependent mode. Moreover, the mRNA levels of Glutation peroxidase and Glutation reductase genes was significantly increased by 2.13±0.07 (p < 0.001) and 1.22±0.05 (p < 0.05) fold, respectively, following treatment withZnONPs.
Conclusion: According to the results of this investigation, the Glutation peroxidase and Glutation reductase gene expression was the key factors of glutathione system in elimination of increasing reactive oxygen species treated with ZnONPs.
 

Abstract Aim: In the present study, The anti-metastatic and cytotoxicity effects of cerium oxide nanoparticles on two MCF-7 and T47D breast cancer cell lines have been studied.
Material and methods: MCF-7 and T47D breast cell lines and normal HEK293 cells were treated with 6.5 mg, 650μg, 65μg, 650ng, 65 ng of cerium oxide nanoparticles and MTT analysis was performed to investigate the toxicity of nanoparticles. Then, NM23 and KAI-1 genes expression were measured by real-time PCR method.
Results: Concentrations of 650 μg and 6.5 mg of CNP resulted in approximately 60% death of MCF-7 and T47D cells. Also, in treatment of normal cells with a concentration of 6.5 mg of CNP, Reduced survival of 25% was observed. Gene expression analysis showed that the two anti-metastatic NM23 and KAI-1 genes did not increase expression under  CNP treatment.
Conclusion: According to the results of this study, the cerium oxide nanoparticles have toxic effects on breast cancer cells, However, this effect varies dose and type of cell line dependent. On the other hand, the study of the anti-metastatic expression of genes showed that this nanoparticle is not capable of enhancing the expression of these genes and therefore is ineffective in controlling cell invasion and is likely to exert its anticancer effect through induction of cell death.