Effect of antineoplastic agents on the expression of human telomerase reverse transcriptase beta plus transcript in MCF-7 cells

Objectives: To evaluate the effect of antineoplastic agents on the expression of human telomerase reverse transcriptase (hTERT) splice variants in MCF-7 cells. Design and methods: We have developed a luminometric hybridization assay for hTERT beta plus transcript. MCF-7 cells were isolated before an...

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Bibliographic Details
Published in:Clinical biochemistry 2004-04, Vol.37 (4), p.299-304
Main Authors: Spiropoulou, Tonia, Ferekidou, Lisa, Angelopoulou, Katerina, Stathopoulou, Aliki, Talieri, Maroulio, Lianidou, Evi S
Format: Article
Language:English
Subjects:
Alternative Splicing - drug effects
Antineoplastic agents
Antineoplastic Agents - pharmacology
Cell Line, Tumor
DNA Primers - genetics
DNA-Binding Proteins
Gene Expression Regulation, Neoplastic - drug effects
hTERT
Humans
Hybridization, Genetic
Luminometric hybridization assay
MCF-7
Polymerase Chain Reaction
RNA, Messenger - genetics
RNA, Messenger - metabolism
RNA, Neoplasm - genetics
RNA, Neoplasm - metabolism
Splice variants
Telomerase - genetics
Transcription, Genetic - drug effects
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Summary:Objectives: To evaluate the effect of antineoplastic agents on the expression of human telomerase reverse transcriptase (hTERT) splice variants in MCF-7 cells. Design and methods: We have developed a luminometric hybridization assay for hTERT beta plus transcript. MCF-7 cells were isolated before and after treatment with antineoplastic agents. A combination of nested RT-PCR and the developed luminometric hybridization assay was used for the specific detection of hTERT beta plus transcript in treated and untreated MCF-7 cells. Amplification of all hTERT splicing variants by nested PCR in the same samples was also performed. Results: MCF-7 cells treated with taxol and etoposide were found positive for all hTERT splicing variants, while the expression of hTERT beta plus transcript did not differ significantly before and after exposure. MCF-7 cells treated with doxorubicin and 5-fluorouracil did not express any of hTERT splicing variants. In the presence of cisplatin, three splicing variants of hTERT were detected. Conclusions: The developed hybridization assay is highly sensitive and specific for the detection of hTERT beta plus transcript in clinical samples.
ISSN:0009-9120
1873-2933
DOI:10.1016/j.clinbiochem.2003.12.008