Background: Age-related cataract (ARC) is a primary cause of visual blindness worldwide. Lens epithelial cell (LEC) apoptosis, in which Fas plays an essential role, is a vital cytological basis for cataractogenesis. However, the regulatory mechanism of Fas-dependent LEC apoptosis is not well understood. This study aimed to investigate whether MicroRNA (miRNA)-124 can regulate LEC apoptosis by targeting polypyrimidine tract-binding protein (PTB) and thereby affecting Fas alternative splicing in ARC. Methods: Lens capsule samples from patients with ARC and cornea donors with a transparent lens were collected. HLE-B3 cells were cultured and treated with hydrogen peroxide (H2 O2 ) to establish an apoptosis model in LECs. The expression of miRNA-124, PTB, Fas, and Fas isoforms in tissues and cell lines was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blotting, polyacrylamide gel electrophoresis, and flow cytometry. The miRNA-124 mimic and inhibitor were transfected into HLE-B3 cells, and the effects of the miRNA-124/PTB/Fas pathway in LECs were assessed by analysis of their related targets. Results: High expression of miRNA-124 and membrane Fas (mFas) mRNA and decreased PTB expression were observed in the lens capsule samples. In cells undergoing H2 O2 -induced apoptosis, mFas expression was increased, accompanied by decreased PTB and increased miRNA-124 expression. Subsequently, miRNA-124 upregulation suppressed PTB expression, elevated the mFas level without affecting total Fas expression and promoted apoptosis; miRNA-124 downregulation exerted the opposite effects. Conclusion: This study revealed that miRNA-124 promotes LEC apoptosis in ARC by upregulating mFas through targeted inhibition of PTB. Moreover, the identification of the miRNA-124/PTB/Fas pathway provides novel insight into Fas-dependent LEC apoptosis.Keywords: Fas; PTB; RNA alternative splicing; age-related cataract; microRNA-124.