Cytotoxicity test with simplified crystal violet staining method using microtitre plates and its application to injection drugs

Saotome, Kyoko; Morita, Hiroshi; Umeda, Makoto

doi:10.1016/0887-2333(89)90039-8

Cited by 238 publications

(149 citation statements)

References 6 publications

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“…To assess the cytotoxicity of venom extract, the crystal violet assay was used to evaluate the optical density based on the amount of dye taken up by the adherent monolayer of cells (Kueng et al 1989;Saotome et al 1989). Briefly, 1 9 10 4 cells were seeded in a volume of 100 ll per well of 96-well flat bottom plates (3 9 10 3 cells/mm 2 ) and allowed to adhere overnight.…”

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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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%

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

2013

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“…Cell morphologies were observed and photographed under a light microscope. Cytotoxicity was evaluated quantitatively by monitoring lactate dehydrogenase (LDH) concentration in culture medium and by crystal violet staining (18,19). LDH is abundant in the cytoplasm and is released into the culture medium accompanying damage of the cell membrane.…”

Section: Cytotoxicity Of Cytokines Against A549 Cellsmentioning

confidence: 99%

Analysis of cytotoxicity induced by proinflammatory cytokines in the human alveolar epithelial cell line A549

et al. 2012

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“…The crystal violet assay is designed to obtain quantitative information about the relative density of adherent cells (20,21). Briefly, after treatment, the medium was removed, 50 μL of 0.5% of crystal violet in 50% methanol was added to each well and incubated for ten minutes at room temperature to stain cell nuclei, and subsequently excess dye was washed out gently by distilled water.…”

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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Cytotoxicity test with simplified crystal violet staining method using microtitre plates and its application to injection drugs

Cited by 238 publications

References 6 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

Analysis of cytotoxicity induced by proinflammatory cytokines in the human alveolar epithelial cell line A549

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

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