Determination of Rates of Dna Synthesis in Cultured Mammalian Cell Populations

Siegers, M. P.; Schaer, Jean‐Claude; Hirsiger, Hans; Schindler, R.

doi:10.1083/jcb.62.2.305

Cited by 14 publications

(4 citation statements)

References 19 publications

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“…The DNA synthesis in cells was determined with the radioactive tracer, 3 H-Me-Thy (thymidine). 20 As shown in Figure 5A (a time study), the DNA synthesis without drug (DMSO control) show an increase of 3 H-Me-Thy incorporation with time. However, in the presence of Tig-S (20ug/ml), the DNA synthesis was inhibited.…”

Section: Inhibition Of Dna Synthesis By Tig-s and Tig-rmentioning

confidence: 83%

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Creation of a novel triterpenoid drug that inhibits DNA synthesis

Chan¹,

Mak²

2021

PPIJ

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Protoaescigenin, an aglycone of the Olean-12-en, was esterified with tigloyl chloride. Three esterification products: Tig-N, with esterification at C-24 position; Tig-R, with esterification at C-24 and C-28 positions and Tig-S, with esterification at C-24 and C-21 were found to have cell-growth inhibition activity with human ovarian cancer cells (ES2). The IC 50 values for Tig-N, Tig-R and Tig-S are 8.6±5.1µg/ml, 3.6±1.7µg/ml and 0.17±0.20µg/ml, respectively. All compounds have a common C-24 esterification, but Tig-S with the both (C-24 and C-21) esterification has the highest activity. Tig-S induces cell-death by the apoptosis and the drug-treated cells were arrested in the S-phase of cell cycles. Treatment with the esterified products of Protoaescigenin inhibits DNA synthesis in K562 cells in dose and time dependent manner. A drug-combination study of Tig-R with three anticancer drugs: Cytarabine, Camptothecin and Daunomycin, indicated that the drug effect of Tig-R is additive to the drug effect of these agents. This is the first report to show that protoaescigenin esterified with tigloyl groups inhibits DNA synthesis and inhibits cell-growth.

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Section: Inhibition Of Dna Synthesis By Tig-s and Tig-rmentioning

confidence: 83%

“…Inhibition of DNA synthesis was determined follows the method described by Siegers et al, 20 in which 3 H-Me-Thy (thymidine) was used as tracer. K562 cells, Human chronic MyelogenousLeukemia (CML), are grown in RPMI-1640.…”

Section: Dna Synthesis Determinationmentioning

confidence: 99%

Creation of a novel triterpenoid drug that inhibits DNA synthesis

Chan¹,

Mak²

2021

PPIJ

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“…Therefore, the cellular synthesis of thymidine was blocked with methotrexate during the labelling period. This procedure ensures that the specific activity of intracellular thymidine triphosphate is equal to the specific activity of thymidine added to the medium and thus constant under all experimental conditions (1,17). Experiments that are not presented here showed that the incorporation of 3H-thymidine was linear with time for at least 4 hours under all conditions used.…”

Section: Regulation Of Cell Growthmentioning

confidence: 99%

Effect of cellular growth on protein metabolism in chicken embryo fibroblasts

Balle

Hendil

1984

Carlsberg Res. Commun.

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Protein synthesis in cultures of chick embryo fibroblasts was measured as the incorporation of ~H-leucine into trichloroacetic acid-insoluble material. Intracellular protein degradation was measured as the release of trichloroacetic acid-soluble radioactivity after pre-labelling cell protein with 3H-leucine.The rate of degradation increased whereas the rates of protein synthesis and cell growth decreased as the cultures approached confluence in growth medium with 1% serum. The rate of protein degradation was reduced when the serum concentration in the culture medium was increased, and the effect of increasing serum concentration was higher in dense than in sparse cultures. There were linear correlations between the rate of protein turnover and culture density, but the slope was lower in medium with 4% chicken serum than in medium with 1% chicken serum.Cell growth was measured as the incorporation of 3H-thymidine into DNA. The correlation between culture density and thymidine incorporation was sigmoid. Density-dependent inhibition of growth was less complete in medium with 4% serum than in 1% serum. The correlations between protein degradation and growth rate were also sigmoid and seemed to differ in media with 1% and 4% serum.

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“…Cell proliferation rate can reveal important information on perturbation of the cell cycle and is used routinely in a variety of biomedical research areas, including oncology and drug discovery. Current cell proliferation assays quantify proliferation either directly by incorporating a modified nucleotide (BrdU/EdU) to the newly synthesized DNA (at S phase of cell cycle) [1][2][3] or indirectly by measuring parameters such as total ATP/DNA content, [4][5][6] metabolic rate, 7,8 substrate impedance changes, [9][10][11][12] etc. Most of the direct techniques are static end point assays.…”

Section: Introductionmentioning

confidence: 99%

Real-time label-free detection of dividing cells by means of lensfree video-microscopy

et al. 2014

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Quantification of cell proliferation and monitoring its kinetics are essential in fields of research such as developmental biology, oncology, etc. Although several proliferation assays exist, monitoring cell proliferation kinetics remains challenging. We present a novel cell proliferation assay based on real-time monitoring of cell culture inside a standard incubator using a lensfree video-microscope, combined with automated detection of single cell divisions over a population of several thousand cells. Since the method is based on direct visualization of dividing cells, it is label-free, continuous, and not sample destructive. Kinetics of cell proliferation can be monitored from a few hours to several days. We compare our method to a standard assay, the EdU proliferation assay, and as proof of principle, we demonstrate concentration-dependent and time-dependent effect of actinomycin D-a cell proliferation inhibitor.

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Determination of Rates of Dna Synthesis in Cultured Mammalian Cell Populations

Cited by 14 publications

References 19 publications

Creation of a novel triterpenoid drug that inhibits DNA synthesis

Creation of a novel triterpenoid drug that inhibits DNA synthesis

Effect of cellular growth on protein metabolism in chicken embryo fibroblasts

Real-time label-free detection of dividing cells by means of lensfree video-microscopy

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