Modification of MTT assay conditions to examine the cytotoxic effects of amitraz on the human lymphoblastoid cell line, WIL2NS

Young, Fiona; Phungtamdet, Wichaya; Sanderson, Barbara J.S.

doi:10.1016/j.tiv.2005.05.001

Cited by 95 publications

(79 citation statements)

References 30 publications

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“…Cells were then treated with venom extract for 24 h. Media and dead cells were washed away after treatment and the remaining cells were stained with crystal violet. The cells were stained with crystal violet for 10 min (Young et al 2005). The microplates were rinsed with distilled water and distained with acetic acid.…”

Section: Cell Viability Testmentioning

confidence: 99%

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The effect of sea anemone (H. magnifica) venom on two human breast cancer lines: death by apoptosis

2013

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Venom from the sea anemone, Heteractis magnifica, has multiple biological effects including, cytotoxic, cytolytic and hemolytic activities. In this study, cytotoxicity induced by H. magnifica venom was investigated using the crystal violet assay on human breast cancer T47D and MCF7 cell lines and normal human breast 184B5 cell line. Apoptosis was also assayed via Annexin V-flourescein isothiocyanate and propidium iodide (PI) staining followed by flow cytometric analysis. Cell cycle progression and mitochondria membrane potential were studied via flow cytometry following PI and JC-1 staining respectively. H. magnifica venom induced significant reductions in viable cell numbers and increases in apoptosis in T47D and MCF7 in dose-dependent manners. A significant apoptosisrelated increase in the sub G1 peak of the cell cycle in both breast cancer cell lines was also observed. Moreover, treatment by venom cleaved caspase-8, caspase-9, and activated caspase-3. Overall, H. magnifica venom was highly cytotoxic to T47D and MCF7 human breast cancer cells, and the phenomenon could be the killing phenomenon via the death receptor-mediated and the mitochondria-mediated apoptotic pathways. Consequently, H. magnifica venom has potential for the development of a breast cancer therapeutic.

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Section: Cell Viability Testmentioning

confidence: 99%

“…OD was measured at 570 nm. The amount of purple dye retained is proportional to the number of cells remaining adhered to the well (Young et al 2005). …”

Section: Cell Viability Testmentioning

confidence: 99%

The effect of sea anemone (H. magnifica) venom on two human breast cancer lines: death by apoptosis

2013

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“…The cytotoxicity effects of crude extracts were determined using the MTT assay (18)(19). Cells were seeded at 10 4 into each well in 100 µL in quadruplicate wells of 96-well flat-bottom plates and incubated for 24 hours (37°C, 5% CO 2 ) to allow attachment of cells.…”

Section: Bioassaysmentioning

confidence: 99%

“…Absorbance values from the MTT and crystal violet assays were converted to cells/well using a standard curve run with each experiment (19). Results from the cell viability assays were expressed as percentages of untreated controls.…”

Section: Bioassaysmentioning

confidence: 99%

Differential susceptibilities of human lung, breast and skin cancer cell lines to killing by five sea anemone venoms

Ramezanpour¹,

Silva²,

Sanderson³

2012

J. Venom. Anim. Toxins incl. Trop. Dis

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Although sea anemones are well known for being rich sources of toxins, including cytolysins and neurotoxins, their venoms and toxins have been poorly studied. In the present study, the venoms from five sea anemones (Heteractis crispa, Heteractis magnifica, Heteractis malu, Cryptodendrum adhaesivum and Entacmaea quadricolor) were obtained by the milking technique, and the potential of these venoms to kill cancer cells was tested on three cell lines (A549 lung cancer, T47D breast cancer and A431 skin cancer). The total protein level in the crude extract was determined by the bicinchoninic acid (BCA) protein assay. The cytotoxicity on different cell lines was assayed using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay which measures survival based on the detection of mitochondrial activity and by the crystal violet assay, which measures survival based on the ability of cells to remain adherent to microplates. The results indicate that the sea anemone venom is cytotoxic to human cancer cells. The A549 cell line was the most sensitive of the cell lines tested with a significant reduction in viability observed at 40 µg/mL. H. malu, C. adhaesivum and E. quadricolor had a significant inhibitory effect on A431 cells. Furthermore, H. malu and C. adhaesivum had a significant inhibitory effect on T47D cell line at 40 µg/mL. In conclusion, the sea anemone venoms tested have the potential to be developed as anticancer agents.

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“…The toxicity of SiO 2 and NDMA was determined by MTT assay as described in literature [22,23]. 1x10 4 cells were seeded in volume of 100 µl into 96-well flat bottom plate.…”

Section: -(45-dimethylthiazol-2-yl)-25-diphenyltetrazolium Bromidementioning

confidence: 99%

Toxicity of N-Nitrosodimethylamine and SiO2 nanoparticles to HaCaT and caco-2 cell lines found in waste water treatment

Almutary¹,

Sanderson²

2017

J Toxicol Health

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Nanoparticles (NPs) offer the possibility of safe removal of pollutants and microbes in water treatment and purification. Nowadays, NPs are used in the detection and purification of water from chemicals and biological substance such as metals (cadmium, copper, lead and zinc) nutrients (nitrate, nitrite ammonia and phosphate) bacteria, viruses, parasite and antibiotics. Metal containing NPs are among the four classes of particles used commonly in water treatment. Membrane technologies such as Ultrafiltration (UF), Nanofiltration (NF) and Reverse Osmosis (RO) have been widely used all over the world especially for water treatment and desalination. NPs after have been incorporated into membranes gained attention due to its ability to enhance membrane permeability, mechanical properties and selectivity in some cases. However, these membranes are suspected to fouling causing NPs and other contaminate to reach waterways. In this study, we tested the toxicity of SiO 2 NPs synthesised by Stöber method and N-Nitrosodimethylamine (NDMA) as potent known carcinogens forms during chlorination on HaCaT and caco-2 cell lines for 4, 24 and 48h using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays also the morphology and size of SiO 2 determined by Scanning electron microscopy (SEM). After exposure to SiO 2 NPs (concentrations between 0.05-2 mg/mL), the concentration of 2 mg/mL inactivated LDH in both cell lines; however, it did not reduce the metabolic activity of both cell lines when MTT assay used. SEM revealed spherical and uniformed SiO 2 particles with size 200 nm in diameter. NDMA (concentrations between 0.1-1000 µg/mL) inactivated LDH leakage in HaCaT and caco-2 also reduced the metabolic activity of HaCaT cell line at 48 hours exposure. The outcome of this study suggest that a concentration of SiO 2 <2 mg/mL used in water treatment can reduce the risk of nanmaterials toxicity to human and possibly the ecosystem. Our results urgefor more studies on the effect of nanomaterials to the aquatic environment and human exposure to NPs.

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Modification of MTT assay conditions to examine the cytotoxic effects of amitraz on the human lymphoblastoid cell line, WIL2NS

Cited by 95 publications

References 30 publications

The effect of sea anemone (H. magnifica) venom on two human breast cancer lines: death by apoptosis

The effect of sea anemone (H. magnifica) venom on two human breast cancer lines: death by apoptosis

Differential susceptibilities of human lung, breast and skin cancer cell lines to killing by five sea anemone venoms

Toxicity of N-Nitrosodimethylamine and SiO2 nanoparticles to HaCaT and caco-2 cell lines found in waste water treatment

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