Effect of antineoplastic agents on the expression of human telomerase reverse transcriptase beta plus transcript in MCF-7 cells.
OBJECTIVE
To evaluate the effect of antineoplastic agents on the expression of human telomerase reverse transcriptase (hTERT) splice variants in MCF-7 cells.


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.
Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, 15771 Athens, Greece.
Tonia
T
Spiropoulou
Lisa
L
Ferekidou
Katerina
K
Angelopoulou
Aliki
A
Stathopoulou
Maroulio
M
Talieri
Evi S
ES
Lianidou
D017398:Alternative Splicing / Q000187:drug effects*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D006824:Hybridization, Genetic; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism*; D019098:Telomerase / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects
D000970:Antineoplastic Agents; D017931:DNA Primers; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D019098:Telomerase