Downregulation of microRNA-181a attenuates hydrogen peroxide-induced human lens epithelial cell apoptosis in vitro.

Molecular Medicine Reports
Zhan ShiPing Liu

Abstract

Apoptosis of human lens epithelial (HLE) cells is a process closely associated with cataract formation. The aim of the present study was to explore the effects of microRNA (miR)‑181a against hydrogen peroxide (H2O2)-induced apoptosis in HLE cells in vitro. The recombinant lentiviral plasmid pLKO. 1‑puro‑miR‑181a was constructed and used to transfect human HLE‑B3 cells with the short hairpin (sh)RNA to silence the expression of miR‑181a. The apoptotic rate of both HLE‑B3 cells in which miR‑181a expression was stably silenced and in untransfected HLE‑B3 cells was assessed in the presence of H2O2 using flow cytometry. The mRNA expression levels of the apoptosis‑related genes caspase-3 (CASP3) and B‑cell lymphoma‑2‑associated X protein (BAX), and of the potential target genes for miR‑181a, c‑MET, cyclooxygenase 2 (COX‑2) and snail family transcriptional repressor 2 (SNAI2) were measured using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) levels were assessed using ELISA. RT‑qPCR analysis revealed that miR‑181a expression was downregulated in HLE‑B3 cells following transfection with miR‑181a‑shRNA. Treatment with H2O2 significantly reduced...Continue Reading

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Apoptosis

Apoptosis is a specific process that leads to programmed cell death through the activation of an evolutionary conserved intracellular pathway leading to pathognomic cellular changes distinct from cellular necrosis