Apoptosis enzyme-linked immunosorbent assay distinguishes anticancer drugs from toxic chemicals and predicts drug synergism

Oskar S. Frankfurt, Awtar Krishan

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The effects of anticancer drugs and toxic compounds on leukemic cells in culture were evaluated by enzyme-linked-immunosorbent assay (ELISA) based on the detection of apoptotic cells by a monoclonal antibody against single-stranded DNA. The concentrations of 13 anticancer drugs, which increased apoptosis ELISA absorbance, were similar to the concentrations decreasing long-term cell survival. Short-term metabolic tetrazolium-based 3-(4,5-dimethylthiazol-yl)-2,5-diphenyformazan bromide (MTT) assay was significantly less sensitive than apoptosis ELISA and the cell survival assay. In contrast to anticancer drugs, 12 toxic chemicals did not increase apoptosis ELISA absorbance at cytotoxic concentrations. The difference between two groups of compounds by apoptosis ELISA was especially large in cultures treated with twofold of concentrations producing 50% inhibition of cell growth: all anticancer drugs induced intense reaction (mean absorbance 2.0), while none of the toxic chemicals induced apoptosis. The application of apoptosis ELISA to chemosensitivity testing was evaluated by its ability to detect synergism of anticancer drug combinations. Among 66 drug combinations tested, only combination of nitrogen mustard with mithramycin was highly synergistic by the apoptosis ELISA, as defined by apoptosis induction with the combination containing each drug at 50% of effective concentration. This combination was also synergistic in the cell survival assay, producing significant cell kill while each drug alone had no effect on cell survival. This synergism was not detected by MTT assay. We conclude that apoptosis ELISA could be useful for drug development and chemosensitivity assessment as it can distinguish clinically useful anticancer drugs from toxic compounds, is as sensitive as the long-term cell survival assay and can detect anticancer drug synergism by rapid evaluation of apoptosis induction.

Original languageEnglish
Pages (from-to)89-99
Number of pages11
JournalChemico-Biological Interactions
Volume145
Issue number1
DOIs
StatePublished - Mar 6 2003
Externally publishedYes

Fingerprint

Drug Synergism
Immunosorbents
Poisons
Assays
Enzyme-Linked Immunosorbent Assay
Apoptosis
Enzymes
Pharmaceutical Preparations
Cell Survival
Cells
Antineoplastic Combined Chemotherapy Protocols
Drug Combinations
Plicamycin
Mechlorethamine
Single-Stranded DNA
Bromides
Cell growth

Keywords

  • Anticancer drugs
  • Apoptosis
  • Chemosensitivity assay
  • Drug screening
  • Enzyme-linked-immunosorbent assay
  • Monoclonal antibody

ASJC Scopus subject areas

  • Toxicology

Cite this

Apoptosis enzyme-linked immunosorbent assay distinguishes anticancer drugs from toxic chemicals and predicts drug synergism. / Frankfurt, Oskar S.; Krishan, Awtar.

In: Chemico-Biological Interactions, Vol. 145, No. 1, 06.03.2003, p. 89-99.

Research output: Contribution to journalArticle

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