Quantitation of mucin rna by pcr reveals induction of both muc2 and
muc5ac mrna levels by retinoids.
Guzman, Karen, Thomas E. Gray, Joo-Heon Yoon, Paul Nettesheim.
Laboratory of Pulmonary Pathobiology, National Institute of
Environmental Health Sciences, Research Triangle Park, NC 27709
APStracts 3:0177L, 1996.
The polydispersity of most human secretory mucin messages has made
them difficult to detect specifically and quantitatively, impeding
the evaluation of the relative expression of the various mucin genes
and their role in normal and pathological conditions. For this
reason, we developed competitive RT-PCR methods to measure the airway
mucins, MUC2 and MUC5AC. Oligonucleotide pairs were designed that
specifically detect MUC2 and MUC5AC, as demonstrated by the size and
sequence of the PCR product and the expected tissue distribution. The
mucin oligonucleotide primers were used to synthesize internal
competitive standards, called MIMICs. Using this assay, the relative
expression of these messages was analyzed in retinoid-replete or
-deprived cultures of normal human tracheobronchial epithelial (NHTBE)
cells. Retinoid deficiency induces squamous metaplasia in vivo and in
vitro. Consistent with these observations and in contrast to a
previous report, retinoid-deprived cultures produced at least an
order of magnitude less MUC2 and MUC5AC message than retinoid-replete
cultures. In summary, this paper describes methodology that can be
applied to the specific and quantitative measurement of mucin
messages and demonstrates that in NHTBE cells the level of MUC2 and
MUC5AC mRNA is increased by retinoids.
Received 11 June 1996; accepted in final form 2 October 1996.
APS Manuscript Number L172-6.
Article publication pending Am. J. Physiol. (Lung Cell. Mol.
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 5 November 1996