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Modified Annexin V/Propidium Iodide Apoptosis Assay For Accurate Assessment of Cell Death
Published on: April 24, 2011
Apoptosis-based drug screening and detection of selective toxicity to cancer cells
Oskar S Frankfurt1, Awtar Krishan
1Experimental Therapeutics Division, Radiation Oncology Department, University of Miami School of Medicine, Miami, FL 33136, USA. apostain@bellsouth.net
Abstract:
The goal of our study was to determine whether an apoptosis assay used after short-term drug exposure could predict selective toxicity to cancer cells. To this end we compared the effect of eight anticancer drugs and 10 toxic compounds without known antitumor activity in cultures of human breast cancer cells and normal diploid fibroblasts by Apoptosis ELISA and growth inhibition assays. There was an overlap in concentration values of drugs and toxins inhibiting proliferation in cancer cells. In contrast, Apoptosis ELISA clearly distinguished between the two groups of compounds. Anticancer drugs induced apoptosis in cancer cells at 0.0015-0.5 microM, while toxins were effective at much higher concentrations of 8.0-50.0 microM. Moreover, six out of the 10 toxins did not induce apoptosis in cancer cells. The normal:cancer cell (N:C) ratio for growth inhibiting concentrations was in a similar range for anticancer drugs and toxins. The N:C ratio for apoptosis inducing concentrations was 33-200 for anticancer drugs and 1.3-3.0 for toxins. Our data indicate that apoptosis assays could be used to detect selective toxicity of anticancer drugs by determining apoptosis induction in cancer cells or through a comparison of apoptosis-inducing concentrations in normal and cancer cells.
Insights
An apoptosis assay can predict selective anticancer drug toxicity. This method clearly distinguishes anticancer drugs from toxins by measuring apoptosis induction in cancer cells, unlike growth inhibition assays.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Selective toxicity is crucial for effective anticancer drug development.
- Distinguishing anticancer agents from general toxins is challenging using traditional proliferation assays.
Purpose of the Study:
- To evaluate the utility of apoptosis assays in predicting selective anticancer drug toxicity.
- To compare the efficacy of apoptosis assays versus growth inhibition assays in differentiating anticancer drugs from toxins.
Main Methods:
- Utilized Apoptosis ELISA and growth inhibition assays.
- Tested eight anticancer drugs and 10 toxic compounds on human breast cancer cells and normal fibroblasts.
- Determined drug concentrations that inhibit proliferation and induce apoptosis.
Main Results:
- Apoptosis ELISA effectively distinguished anticancer drugs from toxins, with anticancer drugs inducing apoptosis at significantly lower concentrations (0.0015–0.5 µM) than toxins (8.0–50.0 µM).
- Growth inhibition assays showed overlapping concentration ranges for drugs and toxins, failing to predict selectivity.
- The normal:cancer cell (N:C) ratio for apoptosis induction was substantially higher for anticancer drugs (33–200) compared to toxins (1.3–3.0).
Conclusions:
- Apoptosis assays are a reliable method for detecting selective anticancer drug toxicity.
- Measuring apoptosis induction in cancer cells or comparing N:C ratios for apoptosis induction can identify potential anticancer agents.
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