This article is part of the supplement: Second International Symposium on the Molecular Biology of Breast Cancer
Development of a high throughput, molecular diagnostic assay for predicting telomerase activity in breast cancer cell lines and tissues
1 Departments of Pathology
2 Human Genetics
3 Massey Cancer Center, Medical College of Virginia Campus at Virginia Commonwealth University, Richmond, VA, USA
4 Department of Biological Sciences, Texas Tech University, Lubbock, TX
5 Baylor College of Medicine, Houston, TX, USA
Breast Cancer Res 2000, 2(Suppl 1):P6.02 doi:10.1186/bcr120
The electronic version of this article is the complete one and can be found online at:
| Published: | 12 March 2000 |
© 2000 Current Science Ltd
Full text
Telomerase is a cellular enzyme that helps to provide genomic stability in tumor cells by maintaining the integrity of telomeres. Telomerase is an RNA-dependent DNA polymerase that contains a protein component (hTERT) and an associated RNA (hTR), which is used as a template for telomere repeat addition. Telomerase activity, while not detectable in most normal human somatic cells, is associated with approximately 85% of malignant human cancers overall, including over 90% of breast cancers.
We have optimized a novel, quantitative, high-throughput telomerase activity assay using fluorescently labelled primers and Real Time quantitation via the ABI Prism 7700 (a.k.a., the TaqMan). Using established breast cancer cell lines and a subset of breast tumors, we demonstrate that telomerase levels quantitated from the TaqMan-based assay closely correlate with values obtained using the traditional, gel-based telomerase activity assay (TRAP). In addition, we have assessed the levels of both hTERT mRNA and hTR in each of our samples via RT-PCR to determine whether relative amounts or a ratio of the two telomerase components correlate with activity in a given sample. Our ultimate goal is to develop a Real Time, fluorescent RT-PCR assay to simultaneously measure hTERT and hTR messages in breast tumor samples, in an attempt to convert the enzymatic telomerase activity assay into a quantitative nucleic acid test to predict levels of activity in routinely processed clinical specimens.