Effect of CLC-2 on the cytoskeleton in human trabecular meshwork cells

Abstract

The chloride channel protein 2 (CLC-2) is important in maintaining the volume of trabecular meshwork cells by adjusting the outflow of aqueous solutions and maintaining the fluid balance. However, little is known concerning the functions of CLC-2 in the cytoskeleton, specifically in human trabecular meshwork (HTM) cells. In the present study, two CLC-2 specific siRNAs (siRNA1 and siRNA2) that target CLC-2 mRNA were designed. The siRNAs were transfected into the HTM cells and the results showed that siRNA1 in particular decreased the expression of CLC-2 by ~45%. Furthermore, an siRNA1-mediated CLC-2 knockdown significantly reconstructed the actin cytoskeleton and formed cross-linked actin networks. In addition, the downregulation of the expression of CLC-2 was associated with increased TGF-? and Smad2 activities in the HTM cells following 24 h of transfection. In conclusion, these results suggest that CLC-2 knockdown promotes trabecular meshwork cytoskeletal disorders and may activate the TGF-?/Smad signaling pathway. Thus, CLC-2 may be a promising and potential novel therapeutic strategy for combating primary open-angle glaucoma.