Volume 2 Supplement 1

Second International Symposium on the Molecular Biology of Breast Cancer

Open Access

Development of a high throughput, molecular diagnostic assay for predicting telomerase activity in breast cancer cell lines and tissues

  • L Elmore1,
  • A Akalin1,
  • L Gollahon4,
  • G Clarke5,
  • M Grimes1,
  • RT Burks1,
  • A Ferreira-Gonzalez1,
  • C Garrett1, 3 and
  • S Holt1, 2, 3
Breast Cancer Research20002(Suppl 1):P6.02

https://doi.org/10.1186/bcr120

Published: 12 March 2000

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.

Authors’ Affiliations

(1)
Departments of Pathology
(2)
Human Genetics
(3)
Massey Cancer Center, Medical College of Virginia Campus at Virginia Commonwealth University
(4)
Department of Biological Sciences, Texas Tech University
(5)
Baylor College of Medicine

Copyright

© Current Science Ltd 2000

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