Department of Ophthalmology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou City, Henan, China.
Bioengineered. 2021 Dec;12(2):9693-9706. doi: 10.1080/21655979.2021.1987068.
Exosomal microRNAs (miRNAs) have been shown to be involved in the regulation of many disease progression, including proliferative vitreoretinopathy (PVR). However, the roles of exosomal miR-4488 and miR-1273 g-5p in PVR progression have not been demonstrated. Transforming growth factor β2 (TGF-β2)-induced ARPE-19 cells were used to stimulate the epithelial-mesenchymal transition (EMT) of cells. Exosomes derived from TGF-β2-induced ARPE-19 cells were identified by transmission electron microscopy and nanoparticle tracking analysis. The expression levels of miR-4488, miR-1273 g-5p and ATP-binding cassette A4 (ABCA4) were measured by quantitative real-time PCR. The promotion levels of exosomes markers, EMT markers, apoptosis markers and ABCA4 were determined by western blot analysis. The migration, invasion and apoptosis of cells were determined by transwell assay, wound healing assay and flow cytometry. Our data showed that miR-4488 and miR-1273 g-5p were lowly expressed in TGF-β2-induced ARPE-19 cells. Overexpressed exosomal miR-4488 and miR-1273 g-5p could inhibit the EMT, migration, invasion, and promote apoptosis in TGF-β2-induced ARPE-19 cells. In addition, ABCA4 was a target of miR-4488 and miR-1273 g-5p. Overexpressed ABCA4 also could reverse the negatively regulation of exosomal miR-4488 and miR-1273 g-5p on the EMT, migration, and invasion of TGF-β2-induced ARPE-19 cells. In conclusion, our data showed that exosomal miR-4488 and miR-1273 g-5p could inhibit TGF-β2-stimulated EMT in ARPE-19 cells through targeting ABCA4.
外泌体 microRNAs（miRNAs）已被证明参与了许多疾病进展的调控，包括增生性玻璃体视网膜病变（PVR）。然而，外泌体 miR-4488 和 miR-1273 g-5p 在 PVR 进展中的作用尚未得到证实。本研究使用转化生长因子β2（TGF-β2）诱导的 ARPE-19 细胞来刺激细胞的上皮-间充质转化（EMT）。通过透射电子显微镜和纳米颗粒跟踪分析鉴定出由 TGF-β2 诱导的 ARPE-19 细胞衍生的外泌体。通过定量实时 PCR 测量 miR-4488、miR-1273 g-5p 和 ATP 结合盒 A4（ABCA4）的表达水平。通过 Western blot 分析测定外泌体标志物、EMT 标志物、凋亡标志物和 ABCA4 的促进水平。通过 Transwell 测定、划痕愈合测定和流式细胞术测定细胞的迁移、侵袭和凋亡。我们的数据表明，miR-4488 和 miR-1273 g-5p 在 TGF-β2 诱导的 ARPE-19 细胞中表达水平较低。过表达的外泌体 miR-4488 和 miR-1273 g-5p 可抑制 TGF-β2 诱导的 ARPE-19 细胞的 EMT、迁移、侵袭，并促进细胞凋亡。此外，ABCA4 是 miR-4488 和 miR-1273 g-5p 的靶标。过表达 ABCA4 也可以逆转外泌体 miR-4488 和 miR-1273 g-5p 对 TGF-β2 诱导的 ARPE-19 细胞 EMT、迁移和侵袭的负调控作用。总之，我们的数据表明，外泌体 miR-4488 和 miR-1273 g-5p 通过靶向 ABCA4 抑制 TGF-β2 刺激的 ARPE-19 细胞 EMT。
