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Originally published In Press as doi:10.1167/iovs.08-2166 on June 14, 2008
 (Investigative Ophthalmology and Visual Science. 2008;49:4753-4759.)
© 2008 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.08-2166
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Sensitization of RPE Cells by {alpha}B-Crystallin siRNA to SAHA-Induced Stage 1 Apoptosis through Abolishing the Association of {alpha}B-Crystallin with HDAC1 in SC35 Speckles

Seung Jin Noh,1,2 Woo Jin Jeong,2,3 Jee Hyun Rho,2,3 Dong Min Shin,3 Hee Bae Ahn,3 Woo Chan Park,3 Sae Heun Rho,3 Young Hwa Soung,1 Tae Hyun Kim,1 Bong Soo Park,4 and Young Hyun Yoo1

1From the Departments of Anatomy and Cell Biology and 3Ophthalmology, Dong-A University College of Medicine and Medical Science Research Center, Busan, South Korea; and the 4College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University, Busan, South Korea.

PURPOSE. To better understand the mechanism underlying the anti-apoptotic activity of {alpha}B-crystallin in RPE cells.

METHODS. Cells of the human retinal pigment epithelial line ARPE-19 were treated with a histone deacetylase inhibitor (HDACI), suberoylanilide hydroxamic acid (SAHA), with or without {alpha}B-crystallin siRNA. To examine the mechanism underlying the cell death induced in ARPE-19 cells, nuclear staining, flow cytometry, DNA electrophoresis, pulse field gel electrophoresis, Western blot analysis, confocal microscopy, and coimmunoprecipitation assay were undertaken.

RESULTS. The present study demonstrated that an HDACI, SAHA, at the usual doses or the silencing of {alpha}B-crystallin by siRNA alone did not effectively induce apoptosis in ARPE-19 cells. Silencing of {alpha}B-crystallin likely abolishes the anti-apoptotic activity of {alpha}B-crystallin. The data indicated that silencing of {alpha}B-crystallin sensitizes ARPE19 cells to SAHA-induced apoptosis and leads them to stage 1 apoptosis. {alpha}B-Crystallin associates with HDAC1 on SC35 speckles, and silencing of {alpha}B-crystallin abolishes this association, resulting in the induction of apoptosis. The data indicated that the association between {alpha}B-crystallin and HDAC1 on SC35 speckles plays a pivotal role in anti-apoptotic activity.

CONCLUSIONS. Knockout of {alpha}B-crystallin may be a promising new approach to enhance therapeutic potency for proliferative vitreoretinopathy without compromising efficacy.






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Copyright © 2008 by the Association for Research in Vision and Ophthalmology