The MTT cell viability assay for cytotoxicity testing in multidrug-resistant human leukemic cells

Marks, Denese C.; Belov, Larissa; Davey, Mary W.; Davey, Ross A.; Kidman, Antony D.

doi:10.1016/0145-2126(92)90114-m

Cited by 149 publications

(95 citation statements)

References 11 publications

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“…7). An MTT cytotoxicity assay, which relies on the activity of mitochondrial dehydrogenases (29), recorded a 20% drop in cell viability at 1,000 yM GSTM or TM. 1,000 tzM cysteine and 300 /.M cystine both decreased viability by 10% relative to 0 MM concentrations.…”

Section: Resultsmentioning

confidence: 99%

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Effect of gold sodium thiomalate and its thiomalate component on the in vitro expression of endothelial cell adhesion molecules.

Newman¹,

To²,

Robinson³

et al. 1994

J. Clin. Invest.

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Endothelial adhesion molecules play an important role in the tissue recruitment of leukocytes in inflammatory conditions such as rheumatoid arthritis. We have investigated the effect of the antirheumatic drug gold sodium thiomalate on adhesion molecule protein and mRNA expression in cultured human endothelial cells. Gold sodium thiomalate inhibited cytokine (TNF, IL-1, IL-4)-stimulated expression of vascular cell adhesion molecule-i and E-selectin but not intercellular adhesion molecule-i on endothelial cells. Gold sodium thiomalate also suppressed TNF-stimulated increases in vascular cell adhesion molecule-i and E-selectin mRNA levels but had no effect on intercellular adhesion molecule-i mRNA. Thiomalate (mercaptosuccinate), but not gold thioglucose or D-penicillamine, mimics the effect of gold sodium thiomalate at equimolar concentrations. We propose that the inhibition of vascular cell adhesion molecule-i and E-selectin expression by gold sodium thiomalate is due to its thiomalate and not its gold component. Gold sodium thiomalate has a direct effect on endothelial adhesion molecule expression, and this may contribute to its antiinflammatory activity. (J.

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Section: Resultsmentioning

confidence: 99%

“…HUVE cells were incubated for 24 h with TNF and the highest concentration of each test drug as per cell ELISA. Cells were then washed with PBS and incubated 2-3 h with MITT (0.4 mg/ml) in basal medium (29). Cells were washed again with PBS, and formazan crystals were dissolved with 100 Il DMSO.…”

Section: Methodsmentioning

confidence: 99%

Effect of gold sodium thiomalate and its thiomalate component on the in vitro expression of endothelial cell adhesion molecules.

Newman¹,

To²,

Robinson³

et al. 1994

J. Clin. Invest.

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show abstract

“…Culture medium in each well was aspirated and replaced with fresh culture medium containing the variable drug concentrations and allowed to grow for a further 96 h. Five wells were used for controls and each dose level. The cell viability was then determined by the MTT assay 28 with minor modifications. In brief, 100 µl of MTT (2 mg/ml in DPBS) (3-(4,5dimethylthiazol-2-yl) 2,5 diphenyl-tetrazolium bromide) were added and the plates were incubated at 37˚C for 2 h in the dark.…”

Section: Methodsmentioning

confidence: 99%

Cytotoxicity of a natural anthraquinone (Aloin) against human breast cancer cell lines with and without ErbB-2: Topoisomerase II-alpha coamplification

Esmat¹,

Tomasetto²,

Rio³

2006

Cancer Biology & Therapy

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In the present study the cytotoxic activity of aloin, a natural anthracycline from Aloe plant, is reported against two human breast cancer cell lines; without (MCF-7) and with (SKBR-3) erbB-2-topoIIα coamplification. MCF-7cell line was shown to be more sensitive to aloin than SKBR-3 demonstrated by MTT and clonogenic assays, from which IC 50 and 50% ICF values are reported to be 60 µg/ml, respectively, in the former cell line and as high as 150 and 80 µg/ml, respectively, in the latter, which are still far below the maximum tolerated dose of the compound. The effect of aloin is suggested to be brought about by more than one mechanism depending on the dose level and tumor phenotype. This was demonstrated by flow cytometric analysis, fluorescence microscopy and western blot analysis, which revealed that aloin at higher concentrations caused a reduction in the proportion of cells undergoing mitosis by induction of apoptosis, inhibition of topo II α protein expression and downregulation of cyclin B1 protein expression in MCF-7 cell line, whereas erbB-2 protein expression was not affected. Topo IIα protein expression was mildly downregulated in SKBR-3 cell line at higher concentrations only.

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“…The cytotoxicity of each drug was determined using an MTT cell viability assay (Marks et al, 1992). Each determination was in triplicate and all experiments were repeated at least once.…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%

Amplification and expression of the ABC transporters ARA and MRP in a series of multidrug-resistant leukaemia cell sublines

O’Neill

Peters²,

Harvie³

et al. 1998

Br J Cancer

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Summary E1000, the most drug-resistant subline from the E-series (CCRF-CEM/E16 to E1000), has been previously shown to express high mRNA levels from two ABC transporter genes associated with multidrug resistance, ARA and MRP. The expression and amplification of both genes has now been characterized for each member of the E-series of drug-resistant sublines and is reported here. Both ARA [detected by reverse transcriptase polymerase chain reaction (RT-PCR)] and MRP (detected by Northern blot analysis) were expressed at low levels in the sensitive parental CEM cell line. An equivalent level of MRP mRNA expression was detected throughout the CEM, E16, E25 and E50 sublines, and there was increasing expression in the El 00, E200 and El 000 sublines. ARA expression was not detected in the El 6, E25, E50 and E100 sublines but was detected by both RT-PCR and Northern blot analysis in the E200 and E1000 sublines. Southern blot analysis indicated the increased levels of MRP and ARA expression resulted from gene amplification and that MRP was first amplified in the E100 subline and ARA in the E200 subline, suggesting that the two genes were not initially co-amplified. Cytogenetic analysis of E1000 cells demonstrated a large addition to chromosome 1 6p, around the region where the ARA and MRP genes are located. Increased expression of ARA is associated with increased colchicine resistance in the E-series of sublines and combined with MRP may account for their resistance phenotype.

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The MTT cell viability assay for cytotoxicity testing in multidrug-resistant human leukemic cells

Cited by 149 publications

References 11 publications

Effect of gold sodium thiomalate and its thiomalate component on the in vitro expression of endothelial cell adhesion molecules.

Effect of gold sodium thiomalate and its thiomalate component on the in vitro expression of endothelial cell adhesion molecules.

Cytotoxicity of a natural anthraquinone (Aloin) against human breast cancer cell lines with and without ErbB-2: Topoisomerase II-alpha coamplification

Amplification and expression of the ABC transporters ARA and MRP in a series of multidrug-resistant leukaemia cell sublines

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