Technical evaluation of thymidine kinase assay in cytosols from breast cancers. EORTC Receptor Study Group Report.
Pilot retrospective studies have pointed to the prognostic value of thymidine kinase (TK) in breast cancer. We studied the Prolifigen TK-REA assay (Sangtec Medical, Sweden), usually applied to serum, for TK analysis in breast cancer cytosols. Reproducibility was good, provided that small volume pipetting was performed carefully. The TK assay was not influenced by the short-term storage of cytosols in liquid nitrogen or at -80 degrees C. However, some steps appeared critical for good laboratory practice. The TK level was affected by thawing the cytosols more than twice. Tumour storage in liquid nitrogen should be preferred over storage at -80 degrees C. The components of the homogenisation buffer, especially sodium molybdate and KCl can have a marked influence on results. Finally, linearity problems arose with some cytosols. Thus, although assay of TK in cytosols is apparently simple, care must be taken in practice. The TK-REA kit should be standardised before widespread use in breast cancer.
Romain
S
S
Laboratoire d'Oncologie Biologique, Faculté de Médecine Nord, Marseille, France.
Spyratos
F
F
Guirou
O
O
Deytieux
S
S
Chinot
O
O
Martin
P M
PM
D001943:Breast Neoplasms / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D005260:Female; D016591:Histocytological Preparation Techniques; D006801:Humans; D015588:Observer Variation; D000091402:Prohibitins; D015203:Reproducibility of Results; D013937:Thymidine Kinase / Q000032:analysis*; D014021:Tissue Preservation
C000717331:PHB2 protein, human; D000091402:Prohibitins; D013937:Thymidine Kinase