Quantification of hTERT splice variants in melanoma by SYBR green real-time polymerase chain reaction indicates a negative regulatory role for the β deletion variant

Lincz, Lisa F.; Mudge, Lisa-Maree; Scorgie, Fiona E.; Sakoff, Jennette A.; Hamilton, Christopher S.; Seldon, Michael

Title: Quantification of hTERT splice variants in melanoma by SYBR green real-time polymerase chain reaction indicates a negative regulatory role for the β deletion variant
Creator: Lincz, Lisa F.; Mudge, Lisa-Maree; Scorgie, Fiona E.; Sakoff, Jennette A.; Hamilton, Christopher S.; Seldon, Michael
Relation: Neoplasia Vol. 10, Issue 10, p. 1131-1137
Relation: http://www.neoplasia.com/abstract.php?msid=1780
Publisher: Neoplasia Press
Resource Type: journal article
Date: 2008
Description: Telomerase activity is primarily determined by transcriptional regulation of the catalytic subunit, human telomerase reverse transcriptase (hTERT). Several mRNA splice variants for hTERT have been identified but it is not clear if telomerase activity is determined by the absolute or relative levels of full length (functional) and variant hTERT transcripts. We have developed a SYBR green-based reverse transcription–quantitative polymerase chain reaction assay for enumeration of the four common hTERT mRNA variants and correlated these with telomerase activity and telomere length in 24 human melanoma cell lines. All except five of the lines expressed four hTERT transcripts, with an overall significant level of co-occurrence between absolute mRNA levels of full length α+/β+ hTERT and the three splice variants α-/β+, α+/β- and α-/β-. On average, α+/β+ made up the majority (48.1%) of transcripts, followed by α+/β- (44.6%), α-/β- (4.4%) and α-/β- (2.9%). Telomerase activity ranged from 1 to 247 relative telomerase activity (RTA), and correlated most strongly with the absolute amount of α+/β+ (R=0.791, p=0.000004) and the relative amount of α+/β- (R = -0.465, P = 0.022). This study shows that telomerase activity in melanoma cells is best determined by the absolute expression of full length hTERT mRNA, and indicates a role for the hTERT β deletion variant in negative regulation of enzyme activity.
Subject: telomerase activity; human telomerase reverse transcriptase (hTERT); mRNA; melanoma cells; β deletion variant; enzyme activity
Identifier: http://hdl.handle.net/1959.13/43312
Identifier: uon:5432
Identifier: ISSN:1522-8002
Language: eng
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