April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
PEDF RNAi Altered Vimentin and B-Crystallin Expression in Human Lens Epithelium
Author Affiliations & Notes
  • J. Yang
    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
    Ophthalmology, Weill Cornell Medical College, New York, New York
  • L. Luo
    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
  • X. Liu
    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
  • M. I. Rosenblatt
    Ophthalmology, Weill Cornell Medical College, New York, New York
  • B. Qu
    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
  • Y. Liu
    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
  • Y. Liu
    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
  • Footnotes
    Commercial Relationships  J. Yang, None; L. Luo, None; X. Liu, None; M.I. Rosenblatt, None; B. Qu, None; Y. Liu, None; Y. Liu, None.
  • Footnotes
    Support  CNSF grant (30772391)
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 6360. doi:
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      J. Yang, L. Luo, X. Liu, M. I. Rosenblatt, B. Qu, Y. Liu, Y. Liu; PEDF RNAi Altered Vimentin and B-Crystallin Expression in Human Lens Epithelium. Invest. Ophthalmol. Vis. Sci. 2010;51(13):6360.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To study relationship of Pigment epithelium-derived factor (PEDF) expression with the expression of vimentin and αB-crystallin by lens epithelial cells.

Methods: : Lens epithelial cells adhering to anterior capsules taken from young donor eyes aged from 20-35 years were cultured and passaged. We designed siRNA constructs to specifically down regulate the expression of Pigment epithelium-derived factor (PEDF) by these primary lens epithelial cells. Quantitative PCR was used to confirm the down regulation of PEDF RNA expression following infection of lens epithelial cells. In order to determine whether altering the expression of PEDF would effect the expression of vimentin or αB-crystallin, we performed western blotting 48 hours after expression of the PEDF directed siRNA.

Results: : PEDF RNA expression in the human lens epithelial cells was strongly down regulated by the three separate siRNA constructs. Western blotting revealed that the down regulation of PEDF expression resulted in a concomitant decrease in expression of vimentin and an increase in αB-crystallin protein

Conclusions: : Decreased expression of PEDF by primary human lens epithelial cells results in a decrease in the expression of vimentin and the increase of αB-crystallin expression, two proteins critical for maintaining lens clarity.

Keywords: cataract • gene/expression 
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