Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of radiosensitivity

J. Carmichael, W. G. DeGraff, A. F. Gazdar, J. D. Minna, J. B. Mitchell

Research output: Contribution to journalArticle

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Abstract

Radiation survival curves were generated for V79 Chinese hamster and two human lung cancer cell lines (NCI-H460 and NCI-H249) with doubling times of 10, 20, and 85 h, respectively, using a standard clonogenic assay, a dye exclusion assay, and a semiautomated colorimetric assay utilizing a tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylformazan bromide. Comparable results for D0 and extrapolation number (n) were observed for all assays in the lines with doubling times of 10 and 20 h. In these instances the tumor cell lines had undergone seven or more doublings after radiation. For the tumor line (H249) with an 80-h doubling time the D0s were comparable between the assays while the extrapolation number was increased in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylformazan bromide assay, a result probably related to the lower number of doublings (< 4) after radiation. We then tested the ability of the assays to detect radiation protection and sensitization using known agents. We found that cysteamine treatment resulted in radioprotection (by a factor of 8 at 8 Gy) while 5-bromo-2-deoxyuridine incorporation caused enhancement of radiation sensitivity in all three assays. We conclude that, while optimal conditions for each cell line (cell number plated and doubling time) must be established, using characterized tumor cell lines, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylformazan bromide assay could be automated and thus be of great value in screening large numbers of potential radiosensitizers or protectors.

Original languageEnglish (US)
Pages (from-to)943-946
Number of pages4
JournalCancer Research
Volume47
Issue number4
StatePublished - 1987

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Radiation Tolerance
Bromides
Radiation
Tumor Cell Line
Tetrazolium Salts
Cysteamine
Cell Line
Radiation Protection
Bromodeoxyuridine
Cricetulus
Lung Neoplasms
Coloring Agents
Cell Count
Survival
Neoplasms

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Carmichael, J., DeGraff, W. G., Gazdar, A. F., Minna, J. D., & Mitchell, J. B. (1987). Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of radiosensitivity. Cancer Research, 47(4), 943-946.

Evaluation of a tetrazolium-based semiautomated colorimetric assay : Assessment of radiosensitivity. / Carmichael, J.; DeGraff, W. G.; Gazdar, A. F.; Minna, J. D.; Mitchell, J. B.

In: Cancer Research, Vol. 47, No. 4, 1987, p. 943-946.

Research output: Contribution to journalArticle

Carmichael, J, DeGraff, WG, Gazdar, AF, Minna, JD & Mitchell, JB 1987, 'Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of radiosensitivity', Cancer Research, vol. 47, no. 4, pp. 943-946.
Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of radiosensitivity. Cancer Research. 1987;47(4):943-946.
Carmichael, J. ; DeGraff, W. G. ; Gazdar, A. F. ; Minna, J. D. ; Mitchell, J. B. / Evaluation of a tetrazolium-based semiautomated colorimetric assay : Assessment of radiosensitivity. In: Cancer Research. 1987 ; Vol. 47, No. 4. pp. 943-946.
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