Colorimetric chemosensitivity testing using sulforhodamine B.
A colorimetric chemosensitivity test was investigated using sulforhodamine B (SRB), which stains protein synthesized by cells, as an end-point marker. Four cultured cell lines, 9 human tumor xenografts serially transplanted into nude mice, and 14 fresh surgical specimens were subjected to this assay. The optimal conditions for the assay were 3-5 x 10(4) cells per well in a 96-microplate, an SRB concentration of 4%, and an incubation time of more than 10 minutes. When mitomycin C, doxorubicin, cisplatin, and 5-fluorouracil were assessed by the SRB assay, the concentration-effect curves revealed a sharp slope between plateaux at low and high concentrations, suggesting that this assay has an excellent sensitivity which can assess the effect of drugs as "all or none." Although this high sensitivity resulted in good reproducibility of the assay for cultured cell lines, the predictive rate of the SRB assay for the chemosensitivity of human tumor xenografts in vivo was limited to 63.9%. As a result, this SRB assay is thought to be useful for evaluating the chemosensitivity of cultured cells as all or none, since it can assess directly cellular protein synthesis, which is one of the most important parameters of cell renewal, with excellent sensitivity.
Kubota
T
T
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
Takahara
T
T
Nagata
M
M
Furukawa
T
T
Kase
S
S
Tanino
H
H
Ishibiki
K
K
Kitajima
M
M
D000818:Animals; D002945:Cisplatin / Q000627:therapeutic use; D003124:Colorimetry; D004317:Doxorubicin / Q000627:therapeutic use; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005472:Fluorouracil / Q000627:therapeutic use; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008819:Mice, Nude; D016685:Mitomycin / Q000627:therapeutic use; D015203:Reproducibility of Results; D012235:Rhodamines; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D012235:Rhodamines; C022027:lissamine rhodamine B; D016685:Mitomycin; D004317:Doxorubicin; D002945:Cisplatin; D005472:Fluorouracil