Direct determination of intracellular daunorubicin in intact confluent monolayers of AT1 prostate carcinoma cells using a multiwell–multilabel counter
The cytostatic drug daunorubicin exerts its toxic action by intercalating into the DNA. The efficacy of daunorubicin depends on the intracellular amount in the tumor cell. Here we have evaluated the use of a multiwell–multilabel reader for the direct determination of the fluorescent cytostatic drug...
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| Veröffentlicht in: | Analytical biochemistry 2008-10, Vol.381 (1), p.81-85 |
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| creator | Sauvant, C. Thews, O. Wirth, C. Gekle, M. |
| description | The cytostatic drug daunorubicin exerts its toxic action by intercalating into the DNA. The efficacy of daunorubicin depends on the intracellular amount in the tumor cell. Here we have evaluated the use of a multiwell–multilabel reader for the direct determination of the fluorescent cytostatic drug daunorubicin in a prostate carcinoma cell line (AT1 R-3327 Dunning prostate carcinoma cells) grown on 24-well plates. We present evidence that this simple fluorescent parameter is a good measure for the toxicologically relevant amount of the drug intercalated into the DNA and, therefore, is a good predictor for the drug’s cytotoxicity. The amount of cationic cytostatics in a tumor cell is primarily a function of the efflux pump protein p-gycoprotein (pGP). Therefore, it is of great value that the assay is also suitable for the estimation of the multidrug resistance efflux pump (pGP) activity. |
| doi_str_mv | 10.1016/j.ab.2008.06.033 |
| format | Article |
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The efficacy of daunorubicin depends on the intracellular amount in the tumor cell. Here we have evaluated the use of a multiwell–multilabel reader for the direct determination of the fluorescent cytostatic drug daunorubicin in a prostate carcinoma cell line (AT1 R-3327 Dunning prostate carcinoma cells) grown on 24-well plates. We present evidence that this simple fluorescent parameter is a good measure for the toxicologically relevant amount of the drug intercalated into the DNA and, therefore, is a good predictor for the drug’s cytotoxicity. The amount of cationic cytostatics in a tumor cell is primarily a function of the efflux pump protein p-gycoprotein (pGP). 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| subjects | Animals Assay ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism Cell Extracts Cell Line, Tumor Cell Proliferation - drug effects Cellular content Chemistry Techniques, Analytical - instrumentation Chemistry Techniques, Analytical - methods Cytostatics Daunorubicin Daunorubicin - analysis DNA, Neoplasm - metabolism Intracellular Space - drug effects Intracellular Space - metabolism Male Multidrug resistance p-Glycoprotein Prostatic Neoplasms - chemistry Rats Spectrometry, Fluorescence Subcellular Fractions Time Factors Verapamil - pharmacology |
| title | Direct determination of intracellular daunorubicin in intact confluent monolayers of AT1 prostate carcinoma cells using a multiwell–multilabel counter |
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