Comparative study of four extraction procedures for urokinase type plasminogen activator and plasminogen activator inhibitor-1 in breast cancer tissues.
The urokinase type plasminogen activator (u-PA) and the plasminogen activator inhibitor-1 (PAI-1) are among the best second-generation prognostic tissue factors in breast cancer. However, different extraction procedures and assay kits are used in different laboratories. A total of 79 breast tumour tissues stored in liquid nitrogen were analysed in this study. We compared u-PA and PAI-1 levels determined with the American Diagnostics (AD) kit after various extraction procedures. The median cytosolic extraction yield in the presence of 0.4 mol/l KCl, calculated relative to extraction in the presence of 10 ml/l Triton X100 when adapted to standard laboratory working hours (incubation for 2 h instead of 12 h) was 74.4% for u-PA and 85.8% for PAI-1. In addition, the correlations were acceptable. Cytosolic extracts prepared with KCl could permit optimal u-PA and PAI-1 assays while also enabling hormone receptors to be determined with the same specimens. Further studies with clinical data are now necessary to determine the prognostic relevance of this extraction procedure.
Romain
S
S
Laboratoire d'Oncologie Biologique-Biopathologie Tissulaire, Faculté de Médecine Nord, Marseille, France.
Spyratos
F
F
Lainé-Bidron
C
C
Bouchet
C
C
Guirou
O
O
Martin
P M
PM
Oglobine
J
J
Magdelénat
H
H
D001943:Breast Neoplasms / Q000201:enzymology / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D002623:Chemistry Techniques, Analytical / Q000379:methods; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D009363:Neoplasm Proteins; D017395:Plasminogen Activator Inhibitor 1; D011933:Reagent Kits, Diagnostic; D011189:Potassium Chloride; D014568:Urokinase-Type Plasminogen Activator