Analysis of human telomerase reverse transcriptase mRNA (hTERT) expressionin 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 det ection of the human telomerase catalytic subunit human telomerase reverse t ranscriptase (hTERT) mRNA. The LightCycler TeloTAGGG hTERT Quantification K it (Roche Molecular Biochemicals) was designed to be used for the highly se nsitive and quantitative detection of hTERT mRNA relative to the house-keep ing gene porphobilinogen deaminase (PBGD). As a tumor progression model, we investigated 26 myxoid liposarcomas (11 pure myxoid grade I, 15 myxoid/rou nd cell grade II/III) for the hTERT expression level and compared the resul ts of the new method with former measurements performed in silver-stained p olyacrylamide gels. Both methods revealed similar results, with real-time R T-PCR being the more accurate quantification technique, which also saves ti me and material. Elevated hTERT expression (cut-off ratio x 100 at 1.3) was an indicator of round cell components and hence for tumor progression in m yxoid liposarcoma. The new method is capable of differentiating between pur e myxoid and myxoid/round cell liposarcomas for hTERT-expression more accur ately.