Expression profile of telomerase subunits in human pleural mesothelioma.
Using the TRAP assay, telomerase activity was previously detected in over 90% of human pleural mesotheliomas (MMs), but not in mesothelial cell cultures (MCCs), suggesting that telomerase re-activation occurs during multi-step mesothelioma carcinogenesis. The present study determined the expression of the telomerase RNA template (hTERC), the telomerase-associated protein (hTEP1), and the telomerase catalytic sub-unit (hTERT), in 16 pleural MMs and 4 MM-derived cell lines, in two pleural solitary fibrous tumours and in six MCCs. Reverse transcription-polymerase chain reaction analysis revealed that hTERT mRNA expression parallels the activity status documented by the TRAP assay, whereas hTERC and hTEP1 mRNA are commonly expressed in all malignant and non-malignant serosal cells and tissues. Three alternatively spliced hTERT transcripts were detected in all telomerase-positive samples, whereas neither variant could be detected in the MCCs. Detection of the hTERT protein with a commercially available antibody was not successful. These results indicate that hTERT expression is rate-limiting for human telomerase activity and that re-activation, rather than up-regulation, of hTERT expression can play a critical role in MM carcinogenesis. While waiting suitable anti-hTERT antibodies, these results provide information for the design of hTERT mRNA-specific in situ probes to study telomerase in archived pre-malignant serosal lesions.
Dhaene
K
K
Laboratory of Pathology, University of Antwerp (UIA), Department of Medicine, Universiteitsplein 1, B-2610 Wilrijk - Antwerp, Belgium.
Wauters
J
J
Weyn
B
B
Timmermans
J P
JP
van Marck
E
E
D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis / Q000235:genetics; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D004848:Epithelium; D005455:Fluorescent Antibody Technique; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008654:Mesothelioma / Q000201:enzymology*; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D015345:Oligonucleotide Probes; D010997:Pleural Neoplasms / Q000201:enzymology*; D012313:RNA; D012333:RNA, Messenger / Q000032:analysis*; D016601:RNA-Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D009363:Neoplasm Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C104342:TEP1 protein, human; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase