Establishing Cell-Based Reporter Systems for the Analysis of hTERT Expression

摘要: Telomeres are the protective structures at end of eukaryotic chromosomes. Telomerase is a ribonucleoprotein that contains both an RNA and protein component for maintenance telomere length. activity detected in majority malignant tumors, but not normal somatic cells, suggesting telomerase reactivation crucial step cell immortality carcinogenesis. The mechanism how activated during tumorigenesis remains unclear. However, expression human reverse transcriptase (hTERT) gene, which encodes catalytic subunit telomerase, has been shown to be major determining factor. To gain insight into mechanisms regulating hTERT facilitate screening agents affect expression, we have established cell-based systems monitoring expression. We linked promoter two different reporter genes encoding green fluorescence (GFP) secreted alkaline phosphatase (SEAP), respectively. These constructs were then transfected H1299 hTERT-BJ1 cells. Stable clones harboring these DNA isolated. In can monitored through quantification GFP or SEAP on automatic plate reader. Using systems, identified several small molecule compounds telomerase.