Department of Neurology, West China Hospital, Sichuan University, No. 37, Guo Xue Xiang, Chengdu City, Sichuan Province 610041, China; Department of Neural Tumor, Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, No. 181, Hanyu Road, Chongqing City 400030, China.
Department of Neurology, West China Hospital, Sichuan University, No. 37, Guo Xue Xiang, Chengdu City, Sichuan Province 610041, China.
Life Sci. 2021 Feb 15;267:118935. doi: 10.1016/j.lfs.2020.118935. Epub 2020 Dec 31.
Long non-coding RNAs (lncRNAs) have been reported to be involved in regulating epilepsy. The purpose of this study is to investigate the possibly regulatory mechanism of small nucleolar RNA host gene 1 (SNHG1) on epilepsy.
Quantitative real-time PCR was utilized to detect the expression of SNHG1, microRNA (miR)-181a, and B-cell lymphoma-2 (BCL-2). Through an enzyme-linked immunosorbent assay, the levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and cyclooxygenase-2 (COX-2) were determined. The viability and apoptosis of CTX-TNA2 cells were measured using MTT assay and flow cytometry analysis, respectively. Western blot assay was performed to analyze the protein levels of Bcl-2, BCL2-associated X, and Caspase-3. The relationships between miR-181a and SNHG1/BCL-2 were confirmed by the dual-luciferase reporter assay.
SNHG1 expression was down-regulated in EP tissues and kainic acid (KA)-induced CTX-TNA2 cells. The apoptosis and release of inflammatory factors (TNF-α, IL-1β, IL-6, and COX-2) in KA-induced CTX-TNA2 cells were suppressed by SNHG1 overexpression and promoted by miR-181a up-regulation. In addition, we confirmed that SNHG1 targeted miR-181a, whereas BCL-2 was a target gene of miR-181a. Negative correlations between SNHG1 and miR-181a, as well as miR-181a and BCL-2 were exhibited. Both the up-regulation of miR-181a and down-regulation of BCL-2 reversed the inhibiting effects of SNHG1 on apoptosis and inflammatory response of KA-induced CTX-TNA2 cells, and the promoting effect upon cell viability.
SNHG1 alleviated the progression of EP by modulating the miR-181a/BCL-2 axis in vitro, thus SNHG1 could act as a possible therapeutic target for treating EP.
长链非编码 RNA（lncRNA）已被报道参与调节癫痫。本研究旨在探讨小核仁 RNA 宿主基因 1（SNHG1）对癫痫的可能调节机制。
采用实时定量 PCR 检测 SNHG1、microRNA（miR）-181a 和 B 细胞淋巴瘤-2（BCL-2）的表达。通过酶联免疫吸附试验测定肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-1β、IL-6 和环氧化酶-2（COX-2）的水平。采用 MTT assay 和流式细胞术分析分别测定 CTX-TNA2 细胞的活力和凋亡。采用 Western blot assay 分析 Bcl-2、BCL2 相关 X 和 Caspase-3 的蛋白水平。通过双荧光素酶报告基因实验证实 miR-181a 与 SNHG1/BCL-2 之间的关系。
SNHG1 在 EP 组织和海人酸（KA）诱导的 CTX-TNA2 细胞中表达下调。SNHG1 过表达抑制 KA 诱导的 CTX-TNA2 细胞凋亡和炎症因子（TNF-α、IL-1β、IL-6 和 COX-2）释放，而 miR-181a 上调促进凋亡和炎症因子释放。此外，我们证实 SNHG1 靶向 miR-181a，而 BCL-2 是 miR-181a 的靶基因。SNHG1 与 miR-181a 以及 miR-181a 与 BCL-2 之间呈负相关。miR-181a 的上调和 BCL-2 的下调均逆转了 SNHG1 对 KA 诱导的 CTX-TNA2 细胞凋亡和炎症反应的抑制作用，以及对细胞活力的促进作用。
SNHG1 通过在体外调节 miR-181a/BCL-2 轴减轻 EP 的进展，因此 SNHG1 可作为治疗 EP 的潜在治疗靶点。
