J Korean Ophthalmol Soc.  2020 May;61(5):472-481. 10.3341/jkos.2020.61.5.472.

Genes Expressed in Steroid-exposed Lens Epithelial Cells as Revealed by Polymerase Chain Reaction

• Yoo WS
• Seo JS
• Cho YW
• Hah YS
• Chung IY
• Seo SW
• Kim SJ

• KMID: 2500719
• DOI: http://doi.org/10.3341/jkos.2020.61.5.472

Abstract

Purpose
We investigated the expression levels of 84 genes in dexamethasone-exposed human lens epithelial cells using polymerase chain reaction (PCR) array analysis.
Methods
The viability and motility of lens epithelial cells were examined after treatment with dexamethasone at 0.01, 0.1, and 1 mg/mL; Western blot was used to evaluate the expression levels of fibronectin, α-smooth muscle actin (α-SMA), and E-cadherin. After 24, 48, and 72 hours of dexamethasone treatment at 0.1 mg/mL, the expression levels of 84 growth factors were analyzed using PCR array.
Results
Cell viability did not change significantly at dexamethasone levels of 0.01 or 0.1 mg/mL, but decreased markedly at 1 mg/mL; motility increased in a concentration-dependent manner at 0.01 and 0.1 mg/mL. Western blot showed that fibronectin levels increased significantly at all dexamethasone concentrations tested; the α-SMA level increased only at 0.01 mg/mL, and E-cadherin levels decreased significantly at all tested concentrations. PCR showed that the levels of FGF1, FGF2, IL-11, regulators of apoptosis (GDNF, IL-1β, and NRG2), and regulators of cell differentiation (BMP5, FGF1, FGF2, and FGF5) decreased more than twofold, whereas the levels of FGF9 and FGF19 increased more than twofold.
Conclusions
PCR performed after exposure of lens epithelial cells to dexamethasone may identify the genes involved in the development of steroid-induced cataracts.

Keyword

Cataract; Lens proteins; Polymerase chain reaction; Steroids