The miR-130a-5p regulates the proliferation and apoptosis of pulmonary epithelial cells by targeting Foxa2 in lung branching morphogenesis

Abstract

Background: In previous study, we confirmed that the miR-130a-5p/Foxa2 axis regulates fetal lung development in congenital diaphragmatic hernia (CDH). We also showed that miR-130a-5p and Foxa2 in lung tissues were localized to the epithelial cells. However, the biological functions of the miR-130a-5p/Foxa2 axis in pulmonary epithelial cells have yet to be fully elucidated. In the present study, we focused on the effect of the miR-130a-5p/Foxa2 axis on the proliferation and apoptosis of pulmonary epithelial cells and explored the mechanisms underlying these processes. Methods: We investigated the roles of miR-130a-5p and Foxa2 with regards to cell viability and apoptosis, by transfecting miR-130a-5p mimics/inhibitors, and Foxa2 overexpressed lentivirus/siRNA, into BEAS-2B cells. We also conducted the CCK-8 assay, EdU staining, and flow cytometry. Chromatin Immunoprecipitation-sequencing (ChIP-Seq) and ChIP-PCR were applied to study the molecular mechanism of miR-130a-5p/Foxa2 axis affecting the the biological properties of pulmonary epithelial cells. Results: We found that the upregulation of miR-130a-5p suppressed cell proliferation and promoted apoptosis, while the downregulation of miR-130a-5p had the opposite effects. Upregulation of the direct target gene of miR-130a-5p, Foxa2, was able to rescue the effects of miR-130a-5p in Human bronchial epithelial (BEAS-2B) cells. Chip-seq and Chip-PCR assays identified potential transcription factor functions for Foxa2; Gli3 and Eya3 were identified as binding sites for Foxa2, and the potential mechanism required our further study. Conclusions: The results showed that miR-130a-5p/Foxa2 axis played an important role in the epigenetic modification of pulmonary epithelial cells, and our current findings provided a potential mechanism for lung branching morphogenesis.