Department of Ophthalmology, Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China.
Bioengineered. 2022 Mar;13(3):5035-5045. doi: 10.1080/21655979.2022.2032966.
Age-related cataract (ARC) is one of the most common causes of vision loss in aging people. This research analyzed the functions and mechanism of long noncoding RNA KCNQ1 overlapping transcript 1 (KCNQ1OT1) in hydrogen peroxide (HO)-stimulated human lens epithelial cells (SRA01/04 cells) in ARC. SRA01/04 cells were stimulated with 200 µM HO to establish oxidative damage in the ARC model. A MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and flow cytometry analysis were conducted to evaluate cell growth and apoptosis. The relevance between KCNQ1OT1 and microRNA (miR)-124-3p or miR-124-3p and BCL-2-like 11 (BCL2L11) was measured through Starbase and a dual luciferase reporter gene assay. The levels of KCNQ1OT1 and miR-124-3p were assessed via quantitative real-time polymerase chain reaction (qRT-PCR). We observed that KCNQ1OT1 was over-expressed and miR-124-3p was low-expressed in HO-stimulated SRA01/04 cells. KCNQ1OT1 interacted with miR-124-3p and negatively mediated its levels. In addition, KCNQ1OT1-siRNA reversed the effects of HO on SRA01/04 cells, evidenced by enhanced cell viability, inhibited apoptotic cells, promoted Bcl-2 expression, and reduced Bax levels. Nevertheless, these observations were inverted after miR-124-3p inhibitor treatment. Likewise, miR-124-3p mimic had a protective effect on HO-stimulated SRA01/04 cells. Our data suggested that BCL2L11 targeted miR-124-3p directly. In summary, the data indicated that lncRNA KCNQ1OT1 down-regulation protected SRA01/04 cells from oxidative stress stimulated damage via the miR-124-3p/BCL2L11 pathway.
年龄相关性白内障（ARC）是老年人视力下降的最常见原因之一。本研究分析了长链非编码 RNA KCNQ1 重叠转录本 1（KCNQ1OT1）在过氧化氢（HO）刺激的人晶状体上皮细胞（SRA01/04 细胞）中在 ARC 中的功能和机制。用 200µM HO 刺激 SRA01/04 细胞建立 ARC 模型的氧化损伤。通过 MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）测定法和流式细胞术分析评估细胞生长和凋亡。通过 Starbase 和双荧光素酶报告基因测定测量 KCNQ1OT1 与 microRNA（miR）-124-3p 或 miR-124-3p 与 BCL2 样 11（BCL2L11）之间的相关性。通过定量实时聚合酶链反应（qRT-PCR）评估 KCNQ1OT1 和 miR-124-3p 的水平。我们观察到 HO 刺激的 SRA01/04 细胞中 KCNQ1OT1 表达上调，miR-124-3p 表达下调。KCNQ1OT1 与 miR-124-3p 相互作用并负调控其水平。此外，KCNQ1OT1-siRNA 逆转了 HO 对 SRA01/04 细胞的作用，表现为细胞活力增强，凋亡细胞减少，Bcl-2 表达增加，Bax 水平降低。然而，在用 miR-124-3p 抑制剂处理后，观察结果相反。同样，miR-124-3p 模拟物对 HO 刺激的 SRA01/04 细胞具有保护作用。我们的数据表明，BCL2L11 直接靶向 miR-124-3p。总之，数据表明 lncRNA KCNQ1OT1 通过 miR-124-3p/BCL2L11 通路下调可保护 SRA01/04 细胞免受氧化应激刺激损伤。
