Abstract
Background
Posterior capsule opacification occurs mainly due to the remnant lens epithelial cell proliferation and migration after cataract surgery. The purpose of this study was to investigate whether small hairpin RNA (shRNA)-mediated gene silencing of transcription factor forkhead box E3 (FOXE3) can be employed to inhibit the expression of FOXE3 and suppress the growth in lens epithelial cells.
Methods
FOXE3-targeted shRNA was transfected into a human lens epithelial cell line (HLEB-3) using Lipofectamine 2000 reagent. Quantitative PCR was used to confirm the downregulation of FOXE3 mRNA expression following infection of lens epithelial cells, and FOXE3 protein expression levels were evaluated by Western blot analysis and immunofluoresence staining. HLEB-3 cell growth after the transduction was analyzed by cell counting and MTT colorimetric assay. Cell cycle of the HLEB-3 cells was examined by flowcytometric analysis.
Results
Compared with the control groups, both mRNA and protein levels of FOXE3 expression were significantly decreased in shRNA-treated groups, and cytostatic effects were obvious within 48 h after transfection. An increased incidence of G1-phase arrest was identified in FOXE3-shRNA transfected HLEB-3 cells.
Conclusions
shRNA-mediated gene silencing of FOXE3 could significantly inhibit cell growth and induce the G1-phase arrest in HLEB-3 cells. Formation of posterior capsular opacification might be repressed if lens epithelial cell growth ceases after the FOXE3 gene is silenced with molecular biology technology.
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Acknowledgments
This study was supported by the National Natural Science Foundation of China (30600698 and 30901637).
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Ye Wang and Wenfeng Li contributed equally to this work
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Wang, Y., Li, W., Wang, Y. et al. Growth inhibition of human lens epithelial cells by short hairpin RNA in transcription factor forkhead box E3 (FOXE3). Graefes Arch Clin Exp Ophthalmol 250, 999–1007 (2012). https://doi.org/10.1007/s00417-012-1944-5
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DOI: https://doi.org/10.1007/s00417-012-1944-5