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KMID : 0360220200610050472
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Journal of the Korean Ophthalmological Society
2020 Volume.61 No. 5 p.472 ~ p.481
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Genes Expressed in Steroid-exposed Lens Epithelial Cells as Revealed by Polymerase Chain Reaction
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Yoo Woong-Sun
Seo Jin-Seok
Cho Yong-Wun
Hah Young-Sool
Chung In-Young
Seo Seong-Wook
Kim Seong-Jae
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Abstract
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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.
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KEYWORD
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Cataract, Lens proteins, Polymerase chain reaction, Steroids
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