High-sensitivity antitumor drug sensitivity testing.
For antitumor drug sensitivity testing we have been performing the succinic dehydrogenase inhibition test using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). A new tetrazolium salt 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenyl-amino) carbonyl]-2H-tetrazolium hydroxide (XTT) has been synthesized recently, and we have been conducting basic studies using it. We were able to obtain a high degree of sensitivity by adding phenazine methosulfate as a promoter in this assay. In these assays, there was a positive correlation between absorbance and cell count and XTT assay was more sensitive than MTT assay. In XTT assay, the production of formazan increases with reaction time over a protracted period of time, we assessed the possibility of performing assays with fewer cells than MTT was used. The results using cancer cell lines showed that when reacted for a longer time (6 h), it was possible to perform adequate assays using this method with 5,000 cells per well, and it will be useful when the amount of biopsy specimen is limited. We also evaluated the inhibition index of each of the drugs, comparing it with the MTT assay.
Department of Surgery, Tokai Central Hospital, Kakamigahara, Japan.
T
T
Kondo
K
K
Wada
M
M
Kawashima
Y
Y
Sato
M
M
Yamauchi
D002452:Cell Count; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors*; D013778:Tetrazolium Salts; D013844:Thiazoles; D014407:Tumor Cells, Cultured
D007202:Indicators and Reagents; D013778:Tetrazolium Salts; D013844:Thiazoles; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; D013385:Succinate Dehydrogenase; C022616:thiazolyl blue