An in-house method which utilizes 14C-thymidine as a substrate was used to assay deoxythymidine kinase in serum. The method is sensitive enough to detect normal levels of serum deoxythymidine kinase and the assay procedure also enables rapid handling of multiple samples. With a total reaction volume of 60 ul, the enzyme reaction was found to be linear with concentrations for up to 650 U/L of TK activity. On studying serum deoxythymidine kinase (s-TK) activity with incubation time, there was a proportional increase in activity with the length of incubation time. "Within-batch" precision showed a coefficient of variation (CV) of 4.7% for serum with extremely high s-TK levels and a CV of 8.8% for serum with normal s-TK levels. S-TK showed a CV of less than 16.0% in its activity when stored at -8 degrees C and at -20 degrees C. The normal reference range obtained for s-TK activity was 8.6 +/- 7.5 U/L.
The proliferative capacity and continuous survival of cells are highly dependent on telomerase expression and the maintenance of telomere length. For this reason, elevated expression of telomerase has been identified in virtually all cancers, including leukemias; however, it should be noted that expression of telomerase is sometimes observed later in malignant development. This time point of activation is highly dependent on the type of leukemia and its causative factors. Many recent studies in this field have contributed to the elucidation of the mechanisms by which the various forms of leukemias increase telomerase activity. These include the dysregulation of telomerase reverse transcriptase (TERT) at various levels which include transcriptional, post-transcriptional, and post-translational stages. The pathways and biological molecules involved in these processes are also being deciphered with the advent of enabling technologies such as next-generation sequencing (NGS), ribonucleic acid sequencing (RNA-Seq), liquid chromatography-mass spectrometry (LCMS/MS), and many others. It has also been established that TERT possess diagnostic value as most adult cells do not express high levels of telomerase. Indeed, studies have shown that prognosis is not favorable in patients who have leukemias expressing high levels of telomerase. Recent research has indicated that targeting of this gene is able to control the survival of malignant cells and therefore offers a potential treatment for TERT-dependent leukemias. Here we review the mechanisms of hTERT regulation and deliberate their association in malignant states of leukemic cells. Further, we also cover the clinical implications of this gene including its use in diagnostic, prognostic, and therapeutic discoveries.