Cytofluorometric determination of thymidine kinase activity in a mixture of normal and neoplastic cells.

A cytofluorometric assay allowing the measurement of thymidine phosphorylation in single cells had been established (Hengstschläger & Wawra, 1993). This assay enables us to correlate intracellular thymidine kinase (TK) activity with the DNA content of single cells. Enzyme activity levels from neuroblastoma cells and normal fibroblasts derived from the same patient were determined.

Cell cycle regulation of deoxycytidine kinase. Evidence for post-transcriptional control.

Deoxycytidine kinase enzyme activity and deoxycytidine kinase mRNA content were determined at different positions of the cell cycle in permanent cell lines. There was no variation of deoxycytidine kinase activity during the cell cycle in two cell lines, whereas in two other lines the enzyme activity was induced (10- and 15-fold) at the G1/S boundary. In contrast, the level of deoxycytidine kinase mRNA never displayed any cell cycle-dependent changes.

Cytofluorometric assay for the determination of thymidine uptake and phosphorylation in living cells.

Thymidine kinase is a key enzyme for the application of drugs in chemotherapy and for diagnosis. Although of great interest, its regulation during cell cycle and differentiation is difficult to study, as current techniques for isolation of cells in different phases of growth are unsatisfactory. An assay that allows the determination of enzymatic activity in situ in single cells would be much faster than present methods and would elegantly avoid synchronization procedures.