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01.03.2003 | Original Article

Comparison of the sulforhodamine B assay and the clonogenic assay for in vitro chemoradiation studies

verfasst von: Bea Pauwels, Annelies E. C. Korst, Christel M. J. de Pooter, Greet G. O. Pattyn, Hilde A. J. Lambrechts, Marc F. D. Baay, Filip Lardon, Jan B. Vermorken

Erschienen in: Cancer Chemotherapy and Pharmacology | Ausgabe 3/2003

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Abstract

Purpose

Since there is a growing interest in preclinical research on interactions between radiation and cytotoxic agents, this study focused on the development of an alternative to the very laborious clonogenic assay (CA).

Methods

The colorimetric sulforhodamine B (SRB) assay was compared to the clonogenic assay for radiosensitivity testing in two lung cancer cell lines (A549, H292), one colon cancer cell line (HT-29) and one breast cancer cell line (MCF-7). In addition, the combination of the radiosensitizing agent gemcitabine and radiation was investigated with both assays.

Results

The dose-response curves obtained with the SRB assay and the CA were very similar up to 6 Gy. The radiosensitivity parameters (SF₂, α, β, MID and ID₅₀) obtained from the SRB assay and the CA were not significantly different between H292, A549 and MCF-7 cells. The radiation dose-response curves for A549 and H292 cells pretreated with 4 nM gemcitabine for 24 h clearly showed a radiosensitizing effect with both assays. The dose-enhancement factors obtained with the SRB assay and the CA were 1.80 and 1.76, respectively, for A549 cells, and 1.52 and 1.41 for H292 cells.

Conclusions

The SRB assay was shown to be as useful as the more traditional CA for research on chemotherapy/radiotherapy interactions in cell lines with moderate radiosensitivity. This assay will be used for more extensive in vitro research on radiosensitizing compounds in these cell lines.

Banasiak D, Barnetson AR, Odell RA, Mameghan H, Russell PJ (1999) Comparison between clonogenic, MTT and SRB assays for determining radiosensitivity in a panel of human bladder cancer cell lines and a ureteral cell line. Radiat Oncol Invest 7:77–85CrossRef

Barnetson AR, Banasiak D, Fisher RJ, Mameghan H, Ribeiro JCC, Brown K, Brown JL, O' Mara SM, Russell PJ (1999) Heterogeneity of in vitro radiosensitivity in human bladder cancer cells. Radiat Oncol Invest 7:66–76CrossRef

Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB (1987) Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity. Cancer Res 47:943–946PubMed

Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55PubMed

Fertil B, Dertinger H, Courdi A, Malaise EP (1984) Mean inactivation dose: a useful concept for intercomparison of human cell survival curves. Radiat Res 99:73–84PubMed

Fricker SP, Buckley RG (1996) Comparison of two colorimetric assays as cytotoxicity endpoints for an in vitro screen for antitumour agents. Anticancer Res 16:3755–3760PubMed

Griffon G, Merlin J-L, Marchal C (1995) Comparison of sulforhodamine B, tetrazolium and clonogenic assays for in vitro radiosensitivity testing in human ovarian cell lines. Anticancer Drugs 6:115–123PubMed

Haselsberger K, Peterson DC, Thomas DGT, Darling JL (1996) Assay of anticancer drugs in tissue culture: comparison of a tetrazolium-based assay and a protein binding dye assay in short-term cultures derived from human malignant glioma. Anticancer Drugs 7:331–338PubMed

Keepers YP, Pizao PE, Peters GJ, van Ark Otte J, Winograd B, Pinedo HM (1991) Comparison of the sulforhodamine B protein and tetrazolium (MTT) assays for in vitro chemosensitivity testing (published erratum appears in Eur J Cancer 1991;27:1717). Eur J Cancer 27:897–900PubMed

10.

Lagarde P, Gheuens EE, De Pooter CM, De Bruijn EA, van der Heyden S, Chomy F, Van Oosterom AT, Scalliet PG (1995) In vitro effects of piracetam on the radiosensitivity of hypoxic cells (adaptation of MTT assay to hypoxic conditions). Bull Cancer 82:929–938PubMed

11.

Latz D, Fleckenstein K, Eble M, Blatter J, Wannenmacher M, Weber KJ (1998) Radiosensitizing potential of gemcitabine (2′,2′-difluoro-2′-deoxycytidine) within the cell cycle in vitro. Int J Radiat Oncol Biol Phys 41:875–882CrossRefPubMed

12.

Leonard CE, Chan DC, Chou TC, Kumar R, Bunn PA (1996) Paclitaxel enhances in vitro radiosensitivity of squamous carcinoma cell lines of the head and neck. Cancer Res 56:5198–204PubMed

13.

Papazisis KT, Geromichalos GD, Dimitriadis KA, Kortsaris AH (1997) Optimization of the sulforhodamine B colorimetric assay. J Immunol Methods 208:151–158CrossRefPubMed

14.

Price P, McMillan TJ (1990) Use of the tetrazolium assay in measuring the response of human tumor cells to ionizing radiation. Cancer Res 50:1392–1396PubMed

15.

Rubinstein LV, Shoemaker RH, Paull KD, Simon RM, Tosini S, Scudiero DA, Monks A, Boyd MR (1990) Comparison of in vitro anticancer-drug-screening data generated with a tetrazolium assay versus a protein assay against a diverse panel of human tumor cell lines. J Natl Cancer Inst 82:1113–1118PubMed

16.

Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82:1107–1112PubMed

Titel: Comparison of the sulforhodamine B assay and the clonogenic assay for in vitro chemoradiation studies
verfasst von: Bea Pauwels
Annelies E. C. Korst
Christel M. J. de Pooter
Greet G. O. Pattyn
Hilde A. J. Lambrechts
Marc F. D. Baay
Filip Lardon
Jan B. Vermorken
Publikationsdatum: 01.03.2003
Verlag: Springer-Verlag
Erschienen in: Cancer Chemotherapy and Pharmacology / Ausgabe 3/2003
Print ISSN: 0344-5704
Elektronische ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-002-0557-9

Springer Medizin

Comparison of the sulforhodamine B assay and the clonogenic assay for in vitro chemoradiation studies