Department of Ophthalmology, The First Affiliated Hospital of Hainan Medical University, Haikou, 571101, Hainan, People's Republic of China.
Key Laboratory of Emergency and Trauma of Ministry of Education, Department of Emergency Surgery, Key Laboratory of Hainan Trauma and Disaster Rescue, The First Affiliated Hospital, Hainan Medical University, Haikou, 571101, Hainan, People's Republic of China.
Int Ophthalmol. 2024 Sep 4;44(1):363. doi: 10.1007/s10792-024-03295-3.
Epithelial-mesenchymal transition (EMT) is a crucial pathological process that contributes to proliferative vitreoretinopathy (PVR), and research indicates that factors present in the vitreous that target cells play pivotal roles in regulating EMT. Experimental studies have confirmed that rabbit vitreous (RV) promotes EMT in human retinal pigment epithelial (RPE) cells. The long noncoding RNA (lncRNA) MALAT1 has been implicated in EMT in various diseases. Thus, this study aimed to investigate the involvement of lncRNA MALAT1 in vitreous-induced EMT in RPE cells.
MALAT1 was knocked down in ARPE-19 cells by short hairpin RNA (shRNA) transfection. Reverse transcription PCR (RT‒PCR) was used to evaluate MALAT1 expression, and Western blotting analysis was used to measure the expression of EMT-related proteins. Wound-healing, Transwell, and cell contraction assays were conducted to assess cell migration, invasion, and contraction, respectively. Additionally, cell proliferation was assessed using the CCK-8 assay, and cytoskeletal changes were examined by immunofluorescence.
MALAT1 expression was significantly increased in ARPE-19 cells cultured with RV. Silencing MALAT1 effectively suppressed EMT and downregulated the associated factors snail1 and E-cadherin. Furthermore, silencing MALAT1 inhibited the RV-induced migration, invasion, proliferation, and contraction of ARPE-19 cells. Silencing MALAT1 also decreased RV-induced AKT and P53 phosphorylation.
In conclusion, lncRNA MALAT1 participates in regulating vitreous-induced EMT in human RPE cells; these results provide new insight into the pathogenesis of PVR and offer a potential direction for the development of antiproliferative drugs.
上皮-间充质转化（EMT）是增生性玻璃体视网膜病变（PVR）的关键病理过程，研究表明，玻璃体中针对细胞的因子在调节 EMT 中发挥关键作用。实验研究已经证实兔玻璃体（RV）促进人视网膜色素上皮（RPE）细胞的 EMT。长链非编码 RNA（lncRNA）MALAT1 已被涉及到多种疾病中的 EMT。因此，本研究旨在探讨 lncRNA MALAT1 在玻璃体诱导的 RPE 细胞 EMT 中的作用。
通过短发夹 RNA（shRNA）转染敲低 ARPE-19 细胞中的 MALAT1。逆转录 PCR（RT-PCR）用于评估 MALAT1 表达，Western blot 分析用于测量 EMT 相关蛋白的表达。划痕愈合、Transwell 和细胞收缩实验分别用于评估细胞迁移、侵袭和收缩。此外，使用 CCK-8 测定法评估细胞增殖，通过免疫荧光法检测细胞骨架变化。
在培养有 RV 的 ARPE-19 细胞中，MALAT1 的表达显著增加。沉默 MALAT1 可有效抑制 EMT，并下调相关因子 snail1 和 E-cadherin。此外，沉默 MALAT1 抑制了 RV 诱导的 ARPE-19 细胞的迁移、侵袭、增殖和收缩。沉默 MALAT1 还降低了 RV 诱导的 AKT 和 P53 磷酸化。
总之，lncRNA MALAT1 参与调节人 RPE 细胞中玻璃体诱导的 EMT；这些结果为 PVR 的发病机制提供了新的见解，并为开发抗增殖药物提供了潜在的方向。
