Chemosensitivity testing of human lung cancer cell lines using the MTT assay

Carmichael, James; Mitchell, James B.; DeGraff, William; Gamson, Janet; Gazdar, Adi F.; Johnson, Bruce E.; Glatstein, Eli; Minna, John D.

doi:10.1038/bjc.1988.125

Cited by 601 publications

(686 citation statements)

References 16 publications

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“…The MTT assay is based on the spectrophotometric measurement of a blue formazan product formed by reduction of MTT by mitochondrial dehydrogenase in viable cells (Carmichael et al, 1987;Wu et al, 1998b). MTT assay was performed daily as described previously by Wu et al, 1997a. For the e ects of all-trans-RA on SCC-25 cells, SCC-25 cells overexpressing mutant RAR-b2(R269Q) or MSCVpac vector transduced SCC-25 cells were seeded in 96-well plates at a density of 4000 cells/well, and treated with all-trans-RA at concentrations ranging from 10 710 ± 10 76 M. Control cultures were treated with an equal volume of ethanol.…”

Section: Mtt Assaymentioning

confidence: 99%

Modulation of retinoic acid receptor function alters the growth inhibitory response of oral SCC cells to retinoids

Dawson

Soprano

et al. 2000

Oncogene

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Section: Mtt Assaymentioning

confidence: 99%

Modulation of retinoic acid receptor function alters the growth inhibitory response of oral SCC cells to retinoids

Dawson

Soprano

et al. 2000

Oncogene

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“…Cells are incubated for a further 3 h at 378C. MTT is converted into an insoluble purple formazan by cleavage of the tetrazolium ring by mitochondrial dehydrogenase enzymes (Carmichael et al, 1987). After 3 h, media is aspirated and cells lysed with 100 ml DMSO, releasing the purple product from the cells.…”

Section: Lipofection/transfection Of A431 Cellsmentioning

confidence: 99%

Inhibition of the mitogen activated protein (MAP) kinase cascade potentiates cell killing by low dose ionizing radiation in A431 human squamous carcinoma cells

et al. 1998

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The molecular mechanism(s) by which tumor cells survive after exposure to ionizing radiation are not fully understood. Exposure of A431 cells to low doses of radiation (1 Gy) caused prolonged activations of the mitogen activated protein (MAP) kinase and stress activated protein (SAP) kinase pathways, and induced p21 Cip-1/WAF1 via a MAP kinase dependent mechanism. In contrast, higher doses of radiation (6 Gy) caused a much weaker activation of the MAP kinase cascade, but a similar degree of SAP kinase cascade activation. In the presence of MAP kinase blockade by the speci®c MEK1 inhibitor (PD98059) the basal activity of the SAP kinase pathway was enhanced twofold, and the ability of a 1 Gy radiation exposure to activate the SAP kinase pathway was increased *sixfold 60 min after irradiation. In the presence of MAP kinase blockade by PD98059 the ability of a single 1 Gy exposure to cause double stranded DNA breaks (TUNEL assay) was enhanced at least threefold over the following 24 ± 48 h. The increase in DNA damage within 48 h was also mirrored by a similar decrease in A431 cell growth as judged by MTT assays over the next 4 ± 8 days following radiation exposure. This report demonstrates that the MAP kinase cascade is a key cytoprotective pathway in A431 human squamous carcinoma cells which is activated in response to clinically used doses of ionizng radiation. Inhibition of this pathway potentiates the ability of low dose radiation exposure to induce cell death in vitro.

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“…Cell proliferation was measured using MTT reduction in a 96 well tissue culture plate assay (Carmichael et al, 1987). Cells were seeded at 5000/well in 200 ml of GM.…”

Section: Determination Of Cell Population Growth In Culturementioning

confidence: 99%

BPV-4 E8 transforms NIH3T3 cells, up-regulates cyclin A and cyclin A-associated kinase activity and de-regulates expression of the cdk inhibitor p27Kip1

O'Brien

Campo

1998

Oncogene

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The E8 open reading frame of Bovine papillomavirus type 4 (BPV-4) encodes a small (42 amino acid) hydrophobic polypeptide localized to cellular membranes and capable of conferring an anchorage-independent (AI) growth phenotype on primary bovine cells co-transfected with BPV-4 E7 ORF and an activated ras gene. To further study the function of E8 independently of other viral gene products, we have expressed it in the murine ®broblast cell line, NIH3T3. Cells expressing E8 are capable of AI growth and escape growth arrest after serum withdrawal. E8 deregulates cyclin A expression, induces transactivation of the human cyclin A gene promoter and increases endogenous protein levels in cells maintained in short-term suspension culture and in lowserum (LS). Both these culture conditions promote downregulation of cyclin A in control cells. In LS growth conditions E8 permits sustained cyclin A-associated kinase activity but not cyclin E-cdk2 activity. Cyclin A-cdk2 activity and, in part, cyclin A gene expression are regulated by the cdk inhibitor p27 Kip1 . Expression of this cdk inhibitor is also de-regulated in E8 cells, with high levels being detected under all culture conditions tested. These data suggest that the ability of BPV-4 E8 to transform NIH3T3 cells is associated with upregulation of cyclin A-associated kinase activity and de-regulated expression of the cdk inhibitor p27 Kip1 and does not rely on down-regulation of p27 Kip1 expression. Analysis of E8 mutants indicate that the hydrophilic tail' of the molecule (residues 31 ± 42) is required for cell transformation, as assessed by anchorage-independent growth, while a form of E8 with expression restricted to the Endoplasmic Reticulum/cis-Golgi membranes by addition of a`KDEL' retention signal revealed that the sub-cellular localization is an important determinant of E8 biological activity.

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Chemosensitivity testing of human lung cancer cell lines using the MTT assay

Cited by 601 publications

References 16 publications

Modulation of retinoic acid receptor function alters the growth inhibitory response of oral SCC cells to retinoids

Modulation of retinoic acid receptor function alters the growth inhibitory response of oral SCC cells to retinoids

Inhibition of the mitogen activated protein (MAP) kinase cascade potentiates cell killing by low dose ionizing radiation in A431 human squamous carcinoma cells

BPV-4 E8 transforms NIH3T3 cells, up-regulates cyclin A and cyclin A-associated kinase activity and de-regulates expression of the cdk inhibitor p27Kip1

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