Department of Cardiology, Chong Gang General Hospital, Chong Qing, China.
Department of gerontology, Hospital of the University of Electronic Science and Technology of China and Sichuan Provincial people's Hospital, Chengdu, Sichuan Province 610072, China.
Exp Mol Pathol. 2020 Aug;115:104436. doi: 10.1016/j.yexmp.2020.104436. Epub 2020 Mar 30.
Hypoxia may cause diseases in human beings. The up- or down- regulation of lncRNAs were identified to possess the ability to protect the myocardial cells from hypoxia injury. This study explored the role of lncRNA GAS5 in sodium Hydrosulfite Induced Hl-1 Cells in vitro. RT-qPCR was applied to measure the expressions of RNAs whereas the cell viability was detected with CCK-8. Luciferase report assay was performed to validate the binding between lncRNA GAS5 and miR-222-3p. The proteins regarding PI3K/AKT signaling were evaluated through western blot analysis. The results showed that the hypoxia could reduce cell viabilities and enhance the apoptosis. Meanwhile, the PI3K/AKT signaling pathway was suppressed as well. lncRNA GAS5 got expressed higher in hypoxic cells whereas the suppressed GAS5 was able to increase cell viabilities while inhibiting apoptosis. MiR-222-3p was the target gene of lncRNA as determined by the luciferase report assay and its expressions were low in hypoxic cells. The overexpressed miR-222-3p could promote proliferation and inhibit apoptosis. Then, the correlation of GAS5 and miR-222-3p was measured which showed that miR-222-3p could resume the functions of GAS5 in cell viabilities and apoptosis through protein regulation in PI3K and AKT signaling pathways.
缺氧可能会导致人类患病。研究发现,lncRNAs 的上调或下调表达具有保护心肌细胞免受缺氧损伤的能力。本研究探讨了长链非编码 RNA GAS5 在亚硫酸氢钠诱导的 HL-1 细胞体外缺氧中的作用。采用 RT-qPCR 测量 RNA 的表达,用 CCK-8 检测细胞活力。通过荧光素酶报告实验验证 lncRNA GAS5 与 miR-222-3p 的结合。通过 Western blot 分析评估与 PI3K/AKT 信号通路相关的蛋白质。结果表明,缺氧可降低细胞活力并增强细胞凋亡,同时抑制 PI3K/AKT 信号通路。在缺氧细胞中,lncRNA GAS5 的表达升高,而抑制 GAS5 能够增加细胞活力并抑制细胞凋亡。荧光素酶报告实验确定 miR-222-3p 是 lncRNA 的靶基因,并且在缺氧细胞中的表达较低。过表达的 miR-222-3p 可促进增殖并抑制凋亡。然后,测量了 GAS5 和 miR-222-3p 之间的相关性,结果表明 miR-222-3p 可以通过 PI3K 和 AKT 信号通路中的蛋白调节恢复 GAS5 在细胞活力和细胞凋亡中的功能。