Eye Institute, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China.
The Department of Ophthalmology, Fengcheng Hospital, Fengxian District, Shanghai, China.
Invest Ophthalmol Vis Sci. 2019 Nov 1;60(14):4670-4680. doi: 10.1167/iovs.19-27211.
Long noncoding RNAs (lncRNAs) are important in disease progression and cellular functions. This study aimed to conduct global lncRNA profiling and characterize the role of lncRNA 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase delta 3-sence RNA 1 (PLCD3-OT1) in the progression of age-related cataract (ARC).
We performed lncRNA expression profiling of lens capsule from ARC groups and age-matched groups using high-throughput RNA-sequencing. Real-time PCR was conducted to detect the expression pattern of lncRNA and mRNA in the clinical samples and cell model. Assays of cell-counting kit-8, 5'-ethynyl-2'-deoxyuridine, TUNEL, and propidium iodide staining were used to detect cell viability, proliferation, apoptosis, and cell cycle. We also performed fluorescence in situ hybridization assay to detect the location of lncRNA, and verified the endogenous competitive RNA mechanism between miRNAs, lncRNAs, and target genes via double-luciferase reporter analyses.
The expression of lncRNA PLCD3-OT1 and PLCD3 were significantly decreased in ARC. PLCD3-OT1 overexpression promoted the expression of PLCD3, cell viability, proliferation, and inhibited cell apoptosis upon oxidative stress, while knockdown of PLCD3 showed the opposite results. Mechanistically, PLCD3-OT1functions through positively regulation the expression of PLCD3. In addition, PLCD3-OT1 may act as a ceRNA to regulate the expression of PLCD3 through competition for miR-224-5p.
PLCD3-OT1 and PLCD3 may become potential therapeutic targets for the prognosis, diagnosis, and treatment of ARC.
长链非编码 RNA（lncRNA）在疾病进展和细胞功能中具有重要作用。本研究旨在进行全局 lncRNA 谱分析，并研究 lncRNA 1-磷酸肌醇-4,5-二磷酸 3-反义 RNA 1（PLCD3-OT1）在年龄相关性白内障（ARC）进展中的作用。
我们使用高通量 RNA 测序对 ARC 组和年龄匹配组的晶状体囊进行 lncRNA 表达谱分析。实时 PCR 检测临床样本和细胞模型中 lncRNA 和 mRNA 的表达模式。细胞计数试剂盒-8、5'-乙炔基-2'-脱氧尿苷、TUNEL 和碘化丙啶染色用于检测细胞活力、增殖、凋亡和细胞周期。我们还进行了荧光原位杂交（FISH）检测以检测 lncRNA 的位置，并通过双荧光素酶报告分析验证了 miRNA、lncRNA 和靶基因之间的内源性竞争 RNA 机制。
在 ARC 中，lncRNA PLCD3-OT1 和 PLCD3 的表达显著降低。PLCD3-OT1 过表达促进了 PLCD3 的表达、细胞活力、增殖，并抑制了氧化应激下的细胞凋亡，而 PLCD3 的敲低则表现出相反的结果。机制上，PLCD3-OT1 通过正向调节 PLCD3 的表达起作用。此外，PLCD3-OT1 可能作为 ceRNA 通过与 miR-224-5p 竞争来调节 PLCD3 的表达。
PLCD3-OT1 和 PLCD3 可能成为 ARC 预后、诊断和治疗的潜在治疗靶点。
