lncRNA TUG1 regulates Smac/DIABLO expression by competitively inhibiting miR-29b and modulates the apoptosis of lens epithelial cells in age-related cataracts.
lncRNA TUG1 regulates Smac/DIABLO expression by competitively inhibiting miR-29b and modulates the apoptosis of lens epithelial cells in age-related cataracts.
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DOI：
10.1097/cm9.0000000000002530
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发表时间：
2023-10-05
影响因子：
6.1
通讯作者：
中科院分区：
文献类型：
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作者：
牛磺酸上调基因1（taurine upregulation gene 1，TUG 1）是最早发现的长链非编码RNA（long non-coding RNA，lncRNA）之一，在多种肿瘤中广泛表达。此外，它促进细胞增殖、分化、凋亡和迁移。然而，我们对它在白内障发病机制中的重要性的理解仍然有限。本研究旨在探讨lncRNA TUG 1通过调控microRNA（miR-29 b）/第二个晶状体源性半胱天冬酶激活因子轴（secondarypa-derivedactivatorofCaspases axis）介导年龄相关性白内障（ARC）透镜上皮细胞凋亡的机制，为白内障的非手术治疗提供新的思路。采用实时荧光定量聚合酶链反应检测TUG 1、miR-29 b和Smac mRNA的表达水平。Western blotting和免疫荧光检测Smac蛋白的表达。流式细胞仪检测细胞凋亡率，kit-8细胞计数法检测细胞增殖率。通过病毒载体构建、共转染、细胞核和细胞质分离、荧光素酶报告基因测定和RNA免疫沉淀验证了lncRNA TUG 1、miR-29 b和Smac之间的靶向调节关系。200 μmol/L H2 O2处理后ARC和HLE-B3细胞TUG 1和Smac表达增加，而miR-29 b表达降低。体外细胞实验证实下调TUG 1表达可抑制透镜上皮细胞凋亡。从机制上讲，Smac表达受miR-29 b负调控。TUG 1竞争性抑制miR-29 b的表达，并导致更多的Smac释放。此外，miR-29 b部分逆转了TUG 1对人透镜上皮细胞系细胞的作用。lncRNA TUG 1通过竞争性抑制miR-29 b增加ARC中透镜上皮细胞的Smac表达并促进其凋亡。这一机制是ARC形成的细胞学基础。基于这些结果，lncRNA TUG 1/miR 29 b/Smac轴可能是调节ARC发育的新分子途径。
As one of the early discovered long non-coding RNAs (lncRNA), taurine upregulation gene 1 (TUG1) has been widely expressed in a variety of tumors. Moreover, it promotes cell proliferation, differentiation, apoptosis, and migration. However, our understanding of its importance in the pathogenesis of cataracts remains limited. This study aimed to explore the mechanism by which lncRNA TUG1 mediates lens epithelial cell apoptosis in age-related cataracts (ARC) by regulating the microRNAs (miR-29b)/second mitochondria-derived activator of caspases axis, and to identify more non-surgical strategies for cataract treatment. The messenger RNA expression levels of TUG1, miR-29b, and Smac were detected using quantitative real-time polymerase chain reaction in vivo and in vitro. The expression of the Smac protein was analyzed by Western blotting and immunofluorescence. Flow cytometry and cell counting kit-8 assays were used to detect the cell apoptosis and proliferation rates, respectively. The targeted regulatory relationship between lncRNA TUG1, miR-29b, and Smac was verified by viral vector construction, co-transfection, nuclear and cytoplasmic separation, luciferase reporter assays, and RNA immunoprecipitation. TUG1 and Smac were expressed at high levels in ARC and HLE-B3 cells treated with 200 μmol/L H2O2, whereas miR-29b expression was decreased. In vitro cell experiments confirmed that down-regulation of TUG1 could inhibit the apoptosis of lens epithelial cells. Mechanistically, Smac expression was negatively regulated by miR-29b. TUG1 competitively inhibited miR-29b expression and caused greater release of Smac. In addition, miR-29b partially reversed the effects of TUG1 on human lens epithelial cell line cells. lncRNA TUG1 increases Smac expression and promotes apoptosis of lens epithelial cells in ARC by competitively inhibiting miR-29b. This mechanism is the cytological basis for ARC formation. Based on these results, the lncRNA TUG1/miR29b/Smac axis may be a new molecular pathway that regulates ARC development.
影响因子：
3.7
作者：
Ng H;Smith DJ;Nagley P
通讯作者：
Nagley P