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Robert N. Weinreb, James D. Lindsey; Metalloproteinase Gene Transcription in Human Ciliary Muscle Cells with Latanoprost. Invest. Ophthalmol. Vis. Sci. 2002;43(3):716-722.
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purpose. The present study was undertaken to determine whether treatment
of ciliary muscle cells with the prostaglandin (PG) analogue
latanoprost acid alters transcription of mRNA for matrix
metalloproteinase (MMP)-1, -2, -3, and -9.
methods. Human ciliary smooth muscle cell cultures were grown to confluence and
treated for 24 hours with medium supplemented with latanoprost acid or
vehicle. Total RNA was then isolated, and the expression of mRNAs for
MMP-1, -2, -3, and -9 were determined using Taqman and energy-transfer
real-time PCR analyses. All results were normalized according
to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in
results. Specificity and linearity of each real-time PCR assay were confirmed by
electrophoresis and serial dilution analysis of oligonucleotides
containing the amplicon sequence. Addition of latanoprost acid for 24
hours increased expression of MMP-1 by 3- to 13-fold in three of five
primary ciliary muscle lines. Addition of 8, 40, and 200 nM latanoprost
acid for 24 hours increased MMP-1 mRNA in a dose-dependent manner.
Analysis of cultures exposed to 200 nM latanoprost acid for 4, 6, 12,
or 24 hours revealed an increase and then a decline of MMP-1 mRNA, with
peak expression at 6 to 12 hours after initiation of treatment.
Parallel assessments of RNA from ciliary muscle cultures exposed to
latanoprost acid for 24 hours revealed increased MMP-1, -3, and -9
mRNAs and reduced MMP-2 mRNA, when compared with RNA from
conclusions. Latanoprost acid induced a dose-dependent increase of MMP-1, -3, and -9
gene transcription in cultured human ciliary smooth muscle cells. These
results are consistent with increased MMPs contributing to the
increased uveoscleral outflow facility observed after topical
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