Expression of telomerase subunits in normal and neoplastic prostate epithelial cells isolated by laser capture microdissection

Abstract

Background: Vertebrates have special structures at the ends of their chromosomes, known as telomeres, which may provide the chromosomes with stability and protect them from exonucleolytic degradation. The shortening of telomeric DNA with each cell division may lead to cell cycle arrest and/or apoptosis of a normal human somatic cell. Telomerase, an RNA-dependent DNA polymerase, elongates the 3'-ends of telomeric DNA. Thus, the presence of telomerase activity may reflect a cell's potential immortal state. The telomerase complex is comprised of several subunits. In the current study, the authors describe the use of laser capture microdissection (LCM) to procure pure matched tumor and normal cell populations from histologic sections and to determine the expression of telomerase subunits in these purified samples.

Methods: Pure matched tumor and normal prostate epithelial cells were procured by LCM using fresh frozen tissue samples obtained from patients undergoing radical prostatectomy. RNA was extracted from LCM captured cells, and the subunits for telomerase were assayed by reverse transcriptase-polymerase chain reaction.

Results: In 18 samples that were captured with LCM, only the catalytic subunit of telomerase, or hTERT, was found to be discriminatory between tumor cells (17 of 18 specimens, 94.4%) and nontumor cells (none of 18 specimens). TP1, a protein that has been shown to be associated with telomerase activity, was expressed in 3 of 18 normal cells (16.7%) and 15 of 18 tumor cells (83.3%). The RNA subunit of telomerase, or hTR, was expressed in 10 of 18 normal cells (55.6%) and 18 of 18 tumor cells (100%). There was no apparent correlation between telomerase subunit(s) expression and Gleason sum score.

Conclusions: Molecular analyses of LCM cells from prostate carcinoma patient samples demonstrated transcriptional up-regulation of all telomerase subunits in the prostatic epithelium. However, only the catalytic subunit of telomerase, hTERT, was found to be discriminatory between neoplastic versus normal cells (94.4% vs. 0%). This finding suggests that the hTERT subunit may be a useful marker for the detection of prostate carcinoma and/or a potential target for therapy.