Telomerase activity and telomerase subunits gene expression patterns in neuroblastoma: a molecular and immunohistochemical study establishing prognostic tools for fresh-frozen and paraffin-embedded tissues.
OBJECTIVE
We have recently demonstrated that telomerase activity (TA) is an independent prognostic factor in neuroblastomas. In the present study, the prognostic impact of TA and gene expression of the three major telomerase subunits is evaluated by molecular and immunohistochemical techniques in fresh-frozen and paraffin-embedded tissues.


METHODS
One hundred thirty-three neuroblastomas of all stages were analyzed for TA. The TA levels of 75 neuroblastoma cases were correlated with gene expression of telomerase subunits hTRT, human telomerase RNA (hTR), and telomerase protein 1 (TP1) by quantitative reverse transcriptase polymerase chain reaction (RT-PCR), using an innovative approach on the LightCycler instrument (Roche Diagnostics, Mannheim, Germany). For selected cases, the applicability of RT-PCR and immunohistochemistry for hTRT expression analysis was investigated in paraffin-embedded tissues. TA and subunit expression patterns were correlated with traditional prognostic indicators and disease outcome.


RESULTS
TA was present in a total of 39 (29.3%) of 133 neuroblastomas and in 31 (29.8%) of 104 initial neuroblastomas without cytotoxic pretreatment. TA was significantly correlated with both event-free and overall survival (P <.0001). Furthermore, we found a significant correlation between expression levels of TA and hTRT (P <.0001) as well as hTR (P <.001). Multivariate analysis revealed only TA and tumor stage but not serum lactate dehydrogenase, MYCN amplification, or age at diagnosis as independent prognostic factors.


CONCLUSIONS
The significant correlation with clinical outcome strongly recommends that analysis of TA be incorporated into the clinical investigation of each individual neuroblastoma at the time of diagnosis. Because the mere presence or absence of TA without further quantification is sufficient basis for predicting disease outcome, the telomeric repeat amplification protocol assay could be complemented with but not replaced by analysis of hTRT or hTR expression.
Poremba
C
C
Gerhard-Domagk-Institute of Pathology and Department of Pediatric Hematology and Oncology, Westfälische Wilhelms-University, Münster, Germany.
Scheel
C
C
Hero
B
B
Christiansen
H
H
Schaefer
K L
KL
Nakayama
J i
Ji
Berthold
F
F
Juergens
H
H
Boecker
W
W
Dockhorn-Dworniczak
B
B
D014408:Biomarkers, Tumor / Q000032:analysis; D015152:Blotting, Northern; D002648:Child; D002675:Child, Preschool; D005260:Female; D005629:Frozen Sections; D015870:Gene Expression; D016259:Genes, myc; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D008297:Male; D015999:Multivariate Analysis; D009447:Neuroblastoma / Q000175:diagnosis* / Q000201:enzymology / Q000235:genetics / Q000401:mortality; D016612:Paraffin Embedding; D011379:Prognosis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015996:Survival Rate; D019098:Telomerase / Q000032:analysis* / Q000235:genetics
D014408:Biomarkers, Tumor; D019098:Telomerase