Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1993 May;67(5):1022–1025. doi: 10.1038/bjc.1993.187

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

M Hengstschläger 1, E Wawra 1
PMCID: PMC1968439  PMID: 8388230

Abstract

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. Using this cytofluorometric assay in a mixture of both cell types the neoplastic cells could be distinguished from the normal fibroblasts because of their higher TK level. A human lymphoblastoid cell line was compared with the cell line KG-1, derived from an acute myelogenous leukaemia, in the same way. The increased enzyme activity enabled us to detect KG-1 cells in a mixture with an 10,000-fold excess of Epstein Barr virus transformed lymphocytes.

Full text

PDF
1022

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adler R., McAuslan B. R. Expression of thymidine kinase variants is a function of the replicative state of cells. Cell. 1974 Jun;2(2):113–117. doi: 10.1016/0092-8674(74)90100-7. [DOI] [PubMed] [Google Scholar]
  2. Bardot V., Luccioni C., Lefrançois D., Muleris M., Dutrillaux B. Activity of thymidylate synthetase, thymidine kinase and galactokinase in primary and xenografted human colorectal cancers in relation to their chromosomal patterns. Int J Cancer. 1991 Mar 12;47(5):670–674. doi: 10.1002/ijc.2910470507. [DOI] [PubMed] [Google Scholar]
  3. Bello L. J. Regulation of thymidine kinase synthesis in human cells. Exp Cell Res. 1974 Dec;89(2):263–274. doi: 10.1016/0014-4827(74)90790-3. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Brodeur G. M. Neuroblastoma: clinical significance of genetic abnormalities. Cancer Surv. 1990;9(4):673–688. [PubMed] [Google Scholar]
  6. Gordon H. L., Bardos T. J., Chmielewicz Z. F., Ambrus J. L. Comparative study of the thymidine kinase and thymidylate kinase activities and of the feedbach inhibition of thymidine kinase in normal and neoplastic human tissue. Cancer Res. 1968 Oct;28(10):2068–2077. [PubMed] [Google Scholar]
  7. Gronowitz J. S., Hagberg H., Källander C. F., Simonsson B. The use of serum deoxythymidine kinase as a prognostic marker, and in the monitoring of patients with non-Hodgkin's lymphoma. Br J Cancer. 1983 Apr;47(4):487–495. doi: 10.1038/bjc.1983.78. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hagberg H., Gronowitz S., Killander A., Källander C., Simonsson B., Sundström C., Oberg G. Serum thymidine kinase in acute leukaemia. Br J Cancer. 1984 Apr;49(4):537–540. doi: 10.1038/bjc.1984.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hengstschläger M., Wawra E. Cytofluorometric assay for the determination of thymidine uptake and phosphorylation in living cells. Cytometry. 1993;14(1):39–45. doi: 10.1002/cyto.990140108. [DOI] [PubMed] [Google Scholar]
  10. Herzfeld A., Greengard O. Enzyme activities in human fetal and neoplastic tissues. Cancer. 1980 Nov 1;46(9):2047–2054. doi: 10.1002/1097-0142(19801101)46:9<2047::aid-cncr2820460924>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  11. Källander C. F., Simonsson B., Gronowitz J. S., Nilsson K. Serum deoxythymidine kinase correlates with peripheral lymphocyte thymidine uptake in chronic lymphocytic leukemia. Eur J Haematol. 1987 Apr;38(4):331–337. doi: 10.1111/j.1600-0609.1987.tb00007.x. [DOI] [PubMed] [Google Scholar]
  12. Källander C. F., Simonsson B., Hagberg H., Gronowitz J. S. Serum deoxythymidine kinase gives prognostic information in chronic lymphocytic leukemia. Cancer. 1984 Dec 1;54(11):2450–2455. doi: 10.1002/1097-0142(19841201)54:11<2450::aid-cncr2820541123>3.0.co;2-r. [DOI] [PubMed] [Google Scholar]
  13. O'Neill K., Abram P., Hannigan B., McKenna G. Elevated serum and mononuclear leukocyte thymidine kinase activities in patients with cancer. Ir Med J. 1987 Sep;80(9):264–265. [PubMed] [Google Scholar]
  14. Shows T. B., Alper C. A., Bootsma D., Dorf M., Douglas T., Huisman T., Kit S., Klinger H. P., Kozak C., Lalley P. A. International system for human gene nomenclature (1979) ISGN (1979). Cytogenet Cell Genet. 1979;25(1-4):96–116. doi: 10.1159/000131404. [DOI] [PubMed] [Google Scholar]
  15. Simonsson B., Källander C. F., Brenning G., Killander A., Ahre A., Gronowitz J. S. Evaluation of serum deoxythymidine kinase as a marker in multiple myeloma. Br J Haematol. 1985 Oct;61(2):215–224. doi: 10.1111/j.1365-2141.1985.tb02820.x. [DOI] [PubMed] [Google Scholar]
  16. Wawra E. Microinjection of deoxynucleotides into mouse cells. No evidence that precursors for DNA synthesis are channeled. J Biol Chem. 1988 Jul 15;263(20):9908–9912. [PubMed] [Google Scholar]
  17. Wawra E., Pöckl E., Müllner E., Wintersberger E. Effect of sodium butyrate on induction of cellular and viral DNA syntheses in polyoma virus-infected mouse kidney cells. J Virol. 1981 Jun;38(3):973–981. doi: 10.1128/jvi.38.3.973-981.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES