AIMS: Telomerase is a ribonucleoprotein enzyme that synthesises telomeres after cell division and maintains chromosomal length and stability thus leading to cellular immortalisation. hTERT (human telomerase reverse transcriptase) gene seems to be the rate-limiting determinant of telomerase reactivation. hTERT mRNA expression was reported to correlate with telomerase activity in cell lines and some human tumours. However the correlation between telomerase activity and hTERT mRNA expression has not been previously examined in human breast cancer. The present study aims to quantitatively measure the expression of hTERT mRNA and telomerase activity in human breast cancer and examine the relationship between these parameters. Furthermore the associations with other parameters including estrogen receptor (ER) and progesterone receptor (PgR) status, DNA ploidy, and S-phase fraction (SPF) are also examined. METHODS: RNA was extracted from 18 breast carcinomas and hTERT mRNA expressions were estimated by reverse transcriptase-PCR (RT-PCR) and Taqman methodology. These tumours had already been analysed for ER and PgR status using ligand-binding assays and had had their DNA ploidy and S-phase fractions measured by flow cytometry. Telomerase activity had already been determined by using a modified telomeric repeat and amplification protocol (TRAP) assay. RESULTS: The expression of hTERT mRNA in the breast tumours ranged between 1.3 and 2.7 x 10(7) copy numbers per micro g of cellular RNA (the median value was 2.7x10(5) and the mean was 3.1 x 10(6)). Telomerase activity was between 0 and 246 units of Total Protein Generated (TPG), where one unit of TPG was equal to 600 molecules, of telomerase substrate primers extended by at least three telomeric repeats. The median level of TPG was 60 units and the mean level was 81 units). Telomerase activity was found to significantly correlate with hTERT expression (r(s)=0.51112, P=0.0302). There was no significant correlation between hTERT and other parameters. CONCLUSION: hTERT mRNA expression significantly correlates with telomerase activity in human breast cancer. This is consistent with the hypothesis that hTERT is the catalytic and rate-limiting determinant subunit of the enzyme.
AIMS: Telomerase is a ribonucleoprotein enzyme that synthesises telomeres after cell division and maintains chromosomal length and stability thus leading to cellular immortalisation. hTERT (human telomerase reverse transcriptase) gene seems to be the rate-limiting determinant of telomerase reactivation. hTERT mRNA expression was reported to correlate with telomerase activity in cell lines and some humantumours. However the correlation between telomerase activity and hTERT mRNA expression has not been previously examined in humanbreast cancer. The present study aims to quantitatively measure the expression of hTERT mRNA and telomerase activity in humanbreast cancer and examine the relationship between these parameters. Furthermore the associations with other parameters including estrogen receptor (ER) and progesterone receptor (PgR) status, DNA ploidy, and S-phase fraction (SPF) are also examined. METHODS: RNA was extracted from 18 breast carcinomas and hTERT mRNA expressions were estimated by reverse transcriptase-PCR (RT-PCR) and Taqman methodology. These tumours had already been analysed for ER and PgR status using ligand-binding assays and had had their DNA ploidy and S-phase fractions measured by flow cytometry. Telomerase activity had already been determined by using a modified telomeric repeat and amplification protocol (TRAP) assay. RESULTS: The expression of hTERT mRNA in the breast tumours ranged between 1.3 and 2.7 x 10(7) copy numbers per micro g of cellular RNA (the median value was 2.7x10(5) and the mean was 3.1 x 10(6)). Telomerase activity was between 0 and 246 units of Total Protein Generated (TPG), where one unit of TPG was equal to 600 molecules, of telomerase substrate primers extended by at least three telomeric repeats. The median level of TPG was 60 units and the mean level was 81 units). Telomerase activity was found to significantly correlate with hTERT expression (r(s)=0.51112, P=0.0302). There was no significant correlation between hTERT and other parameters. CONCLUSION:hTERT mRNA expression significantly correlates with telomerase activity in humanbreast cancer. This is consistent with the hypothesis that hTERT is the catalytic and rate-limiting determinant subunit of the enzyme.
Authors: Fariborz Rashid-Kolvear; Michael A S Taboski; Johnny Nguyen; Dong-Yu Wang; Lea A Harrington; Susan J Done Journal: BMC Cancer Date: 2010-07-22 Impact factor: 4.430