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

doi:10.1186/bcr120

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Breast Cancer Res
Volume 2
Suppl 1

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Elmore L
Akalin A
Gollahon L
Clarke G
Grimes M
Burks RT
Ferreira-Gonzalez A
Garrett C
Holt S

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Elmore L
Akalin A
Gollahon L
Clarke G
Grimes M
Burks RT
Ferreira-Gonzalez A
Garrett C
Holt S
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This article is part of the supplement: Second International Symposium on the Molecular Biology of Breast Cancer .

Meeting abstract

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

¹Departments of Pathology

²Human Genetics

³Massey Cancer Center, Medical College of Virginia Campus at Virginia Commonwealth University, Richmond, VA, USA

⁴Department of Biological Sciences, Texas Tech University, Lubbock, TX

⁵Baylor College of Medicine, Houston, TX, USA

from Second International Symposium on the Molecular Biology of Breast Cancer
Lillehammer, Norway. 12–16 March 2000

Breast Cancer Res 2000, 2(Suppl 1):P6.02doi:10.1186/bcr120

Published:

12 March 2000

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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.

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

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