Analysis of human telomerase reverse transcriptase mRNA (hTERT) expression in myxoid liposarcomas using LightCycler real-time quantitative reverse transcriptase-polymerase chain reaction

We describe a convenient, nonradioactive reverse transcription – polymerase chain reaktion (RT‐PCR) method for the rapid and accurate quantitative detection of the human telomerase catalytic subunit human telomerase reverse transcriptase (hTERT) mRNA. The LightCycler TeloTAGGG hTERT Quantification K...

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Bibliographic Details
Published in:Electrophoresis Vol. 22; no. 6; pp. 1098 - 1101
Main Authors: Schneider-Stock, Regine, Emrich, Thomas, Peters, Brigitte, Jaeger, Viola, Roessner, Albert
Format: Journal Article
Language:English
Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.04.2001
Subjects:
Computer Systems
DNA-Binding Proteins
Gene Expression
Human telomerase reverse transcriptase
Humans
LightCycler
Liposarcoma
Liposarcoma, Myxoid - enzymology
Liposarcoma, Myxoid - genetics
Real-time polymerase chain raection
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger - analysis
Telomerase - genetics
ISSN:0173-0835, 1522-2683
Online Access:Get full text
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Summary:We describe a convenient, nonradioactive reverse transcription – polymerase chain reaktion (RT‐PCR) method for the rapid and accurate quantitative detection of the human telomerase catalytic subunit human telomerase reverse transcriptase (hTERT) mRNA. The LightCycler TeloTAGGG hTERT Quantification Kit (Roche Molecular Biochemicals) was designed to be used for the highly sensitive and quantitative detection of hTERT mRNA relative to the house‐keeping gene porphobilinogen deaminase (PBGD). As a tumor progression model, we investigated 26 myxoid liposarcomas (11 pure myxoid grade I, 15 myxoid/round cell grade II/III) for the hTERT expression level and compared the results of the new method with former measurements performed in silver‐stained polyacrylamide gels. Both methods revealed similar results, with real‐time RT‐PCR being the more accurate quantification technique, which also saves time and material. Elevated hTERT expression (cut‐off ratio×100 at 1.3) was an indicator of round cell components and hence for tumor progression in myxoid liposarcoma. The new method is capable of differentiating between pure myxoid and myxoid/round cell liposarcomas for hTERT‐expression more accurately.
Bibliography:ark:/67375/WNG-KRM742ZW-M
ArticleID:ELPS1098
istex:D38BCF78C8855678755522946D7790EAC81A2BB6
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0173-0835
1522-2683
DOI:10.1002/1522-2683()22:6<1098::AID-ELPS1098>3.0.CO;2-T