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High-sensitivity antitumor drug sensitivity testing

T Kondo1, K Wada, M Kawashima

  • 1Department of Surgery, Tokai Central Hospital, Kakamigahara, Japan.

Oncology
|November 1, 1994
PubMed
Summary
This summary is machine-generated.

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The new XTT assay, enhanced with phenazine methosulfate, offers higher sensitivity for antitumor drug testing than the MTT assay. It allows for accurate results with fewer cells, especially beneficial for limited biopsy specimens.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Pharmacology

Background:

  • The succinic dehydrogenase inhibition test using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) is a standard for antitumor drug sensitivity testing.
  • A novel tetrazolium salt, XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenyl-amino) carbonyl]-2H-tetrazolium hydroxide), has been developed.

Purpose of the Study:

  • To evaluate the sensitivity and applicability of the new XTT assay for antitumor drug sensitivity testing.
  • To compare the performance of the XTT assay with the established MTT assay.

Main Methods:

  • Basic studies were conducted using the XTT assay with phenazine methosulfate as a promoter.
  • Correlation between absorbance and cell count was assessed.
  • Sensitivity comparison between XTT and MTT assays was performed.

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  • Assay optimization for reduced cell numbers and extended reaction times (6 hours) was investigated.
  • Inhibition indices of various drugs were evaluated using both XTT and MTT assays.

    • Main Results:

    • The XTT assay demonstrated a positive correlation between absorbance and cell count.
    • The XTT assay showed higher sensitivity compared to the MTT assay.
    • Adequate assays were achievable with as few as 5,000 cells per well when using a 6-hour reaction time.
    • The XTT assay proved effective for drug sensitivity testing with limited cellular material.

    Conclusions:

    • The XTT assay, particularly with phenazine methosulfate, is a highly sensitive method for antitumor drug sensitivity testing.
    • The XTT assay's ability to function effectively with fewer cells and longer reaction times makes it a valuable tool, especially when dealing with limited biopsy specimens.
    • The XTT assay provides a sensitive and potentially more efficient alternative to the MTT assay for evaluating drug responses in cancer cell lines.