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N. Kondo, K. Ishibashi, S. Honda, Y. Tsukahara, A. Negi; Effects of Cryoinjury on ARPE-19: Identification of Gene Expressions Associated With the Pathogenesis of Proliferative Vitreoretinopathy. Invest. Ophthalmol. Vis. Sci. 2007;48(13):5721.
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© ARVO (1962-2015); The Authors (2016-present)
Cryoretinopexy to be often used in the surgery of retinal detachment increases the risk of development of proliferative vitreoretinopathy (PVR). To elucidate relations between cryoretinopexy and the development of PVR, we analyzed the alteration of gene expression profiles of retinal pigment epithelial (RPE) cells treated with a cryoinjury.
The human RPE cell line, ARPE-19, was grown on culture dishes, then serum was withdrawn for 2 days. Culture dishes were put into -80 degree freezer for 20 seconds to treat ARPE-19 cells with the cryoinjury. The temperature on the surface of empty culture dishes were measured with a thermometer preliminarily. Total RNA was extracted from cultures at 2 hours after the treatment. Cultures untreated with the cryoinjury served as controls. Gene expression profiles in both cultures were compared using microarray analysis. Real-time PCR was conducted to confirm results of microarray analysis.
The temperature of culture dishes put into the freezer for 20 seconds fell to -19.4 degrees. Among up-regulated genes detected by microarray analysis in cultures treated with the cryoinjury, PDGF-A, PDGF-B, IL-6, VEGF-A, fibronectin, and thrombospondin-1 were up-regulated 8.2-fold, 5.5-fold, 3.0-fold, 3.9-fold, 2.4-fold, and 2.1-fold in real-time PCR assay, respectively.
This study demonstrated direct effects of cryoinjury on the alteration of gene expression profiles of ARPE-19 cells, which may contribute to the development of PVR in the form of migration and transdifferentiation of RPE cells, inflammation, cell growth, and formation of proliferative membrane.
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