Quantitative immunohistochemical analysis of the glutathione S-transferase GSTM1: in situ phenotyping in archival material.
1. GSTM1 is present in only approximately 50% of Caucasian individuals and deficiency of GSTM1 is associated with susceptibility to a growing number of diseases, especially cancer. Thus, a method that would allow accurate, retrospective determination of the GSTM1 phenotype in different patient populations would have many applications. 2. Developed, therefore, is a quantitative, image-analysis-based immunohistochemical technique for the analysis of GSTM1 protein in paraffin-embedded tissue samples. It was applied to the determination of the GSTM1 phenotype using liver biopsies taken from 70 patients. 3. Of the 70 cases (depending on the cut-off point), 51-54% were deficient in GSTM1. A single 27 kD band characteristic for GSTM1 was found in seven of 16 cases analysed by Western blotting using the same GSTM1 antibody as in the immunohistochemical analysis. There was a good correlation (r = 0.87) between the staining intensity of the GSTM1 band and the staining intensity evaluated by immunohistochemistry. 4. It is concluded that this quantitative immunohistochemical method permits accurate determination of the GSTM1 phenotype and is well suited for retrospective analysis of GSTM1 expression in specific tissues in situ.
Fritz
P
P
Zentrum für Klinische Pathologie, Robert-Bosch-Krankenhaus, Stuttgart, Germany.
Sperker
B
B
Mürdter
T E
TE
Späth
M
M
Schwarzmann
P
P
Kroemer
H K
HK
Eichelbaum
M
M
Kivistö
K T
KT
D000328:Adult; D000368:Aged; D015153:Blotting, Western / Q000379:methods; D003593:Cytoplasm / Q000201:enzymology; D005234:Fatty Liver / Q000201:enzymology; D005260:Female; D005982:Glutathione Transferase / Q000032:analysis*; D006689:Hodgkin Disease / Q000201:enzymology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008099:Liver / Q000201:enzymology*; D008107:Liver Diseases / Q000201:enzymology*; D008297:Male; D008875:Middle Aged; D016612:Paraffin Embedding; D012016:Reference Values; D015203:Reproducibility of Results
D005982:Glutathione Transferase