Exfoliated oral cell messenger RNA: suitability for biomarker studies.
The efficacy of chemoprevention trials can be improved by the use of biomarkers of carcinogenesis that serve as surrogate end points. The aim of this study was to assess the perspectives of using mRNA isolated from oral exfoliated cells for biomarker research in chemoprevention of upper aerodigestive tract cancer. When using reverse transcription-PCR in combination with Southern blotting and hybridization, it was possible to detect transcripts from only five cells. With the quantitative RNase protection assay, we could only detect highly abundant transcripts. The integrity of the RNA was verified by Northern blotting, which showed a variable degree of degradation, depending on the gene studied. Interestingly, although specific transcripts were found to be intact to a certain extent, the rRNA appeared to be completely degraded, suggesting that a specific protein synthesis shut-off mechanism exists in terminally differentiated oral epithelial cells. Altogether, this differential RNA degradation makes accurate measurement of transcript levels of most genes, as determined in exfoliated oral cells, unreliable. Because this RNA degradation process is likely to start before the cells are shed from the tissue, the results of measurements of transcript levels in biopsies of oral tissue should be interpreted with caution.
Klaassen
I
I
Department of Otolaryngology/Head and Neck Surgery, University Hospital Vrije Universiteit, Amsterdam, The Netherlands. p6
Copper
M P
MP
Brakenhoff
R H
RH
Smeets
S J
SJ
Snow
G B
GB
Braakhuis
B J
BJ
D000328:Adult; D014408:Biomarkers, Tumor / Q000235:genetics*; D015152:Blotting, Northern; D017931:DNA Primers; D005260:Female; D006801:Humans; D008297:Male; D009062:Mouth Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D014408:Biomarkers, Tumor; D017931:DNA Primers; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase