Use of monochlorobimane for glutathione measurements in hamster and human tumor cell lines.
The use of monochlorobimane (MCIB) as a fluorescence label for glutathione (GSH) quantitation was investigated in human tumor cell lines. When MCIB was used with a hamster fibroblast cell line under conditions where GSH was either depleted or elevated, an excellent correlation between bimane-GSH fluorescence and the standard cyclic GSH reductase assay (Tietze's) was accomplished. When the MCIB technique was applied to a human lung adenocarcinoma cell line, little or no GSH labeling was noted even at MCIB levels 10X higher than that used for the hamster line. HPLC analysis suggested that the source of the problem may be the affinity for MCIB to glutathione S-transferase. By using higher dye concentrations and longer staining times, adequate staining was possible. While the MCIB technique may have problems quantitating GSH levels between cell types, the possibility of examining GSH heterogeneity in solid tumor biopsies remains feasible.
Cook
J A
JA
Radiation Oncology Branch, National Cancer Institute, Bethesda, MD 20892.
Pass
H I
HI
Russo
A
A
Iype
S
S
Mitchell
J B
JB
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D005347:Fibroblasts / Q000032:analysis; D005434:Flow Cytometry; D005978:Glutathione / Q000032:analysis*; D006801:Humans; D009374:Neoplasms, Experimental / Q000032:analysis*; D011720:Pyrazoles; D014407:Tumor Cells, Cultured
D011720:Pyrazoles; C059597:monochlorobimane; D005978:Glutathione