Department of Orthopaedic, The Third Xiangya Hospital of Central South University, Changsha, 410008, Hunan, China.
Department of Orthopaedic, Xiangya Hospital Central South University, Changsha, 410008, Hunan, China.
Neurochem Res. 2021 Apr;46(4):945-956. doi: 10.1007/s11064-020-03224-7. Epub 2021 Jan 30.
Spinal cord injury (SCI) is a serious neurological disease. Long non-coding RNA (lncRNA) small nucleolar RNA host gene (SNHG1) and microRNA-362-3p (miR-362-3p) were confirmed to be related to neurological disorders. However, it is unclear whether SNHG1 was involved in the development of SCI via regulating miR-362-3p. PC12 cells were treated with lipopolysaccharide (LPS) to imitate the in vitro cell model of SCI. Cell ciability and apoptosis rate were detected by cell counting kit-8 (CCK-8) assay and flow cytometry assay. The levels of SNHG1, miR-362-3p, and Janus kinase-2 (Jak2) were examined by quantitative real-time polymerase chain reaction (qRT-PCR). The dual-luciferase reporter assay, RNA pull-down assay, and RNA immunoprecipitation (RIP) assay were performed to verify the interaction between miR-362-3p and SNHG1 or Jak2. Besides, the levels of apoptosis- and autophagy- related proteins were detected by western blot assay. In present research, LPS suppressed cell viability, and induced apoptosis and autophagy in PC12 cells. SNHG1 knockdown could affect cell viability, and suppress cell apoptosis and autophagy in LPS-treated PC12 cells. Moreover, miR-362-3p was a target of SNHG1, miR-362-3p targeted Jak2 and negatively regulated Jak2/stat3 pathway. Our data also demonstrated that SNHG1 depletion inactivated Jak2/stat3 pathway to affect cell viability and confine apoptosis, autophagy in LPS-treated PC12 cells. Taken together, SNHG1 regulated cell viability, apoptosis and autophagy in LPS-treated PC12 cells by activating Jak2/stat3 pathway via sponging miR-362-3p.
脊髓损伤(SCI)是一种严重的神经系统疾病。长链非编码 RNA(lncRNA)小核仁 RNA 宿主基因(SNHG1)和 microRNA-362-3p(miR-362-3p)已被证实与神经紊乱有关。然而,SNHG1 是否通过调节 miR-362-3p 参与 SCI 的发展尚不清楚。用脂多糖(LPS)处理 PC12 细胞,模拟 SCI 的体外细胞模型。用细胞计数试剂盒-8(CCK-8)法和流式细胞术检测细胞活力和细胞凋亡率。用实时定量聚合酶链反应(qRT-PCR)检测 SNHG1、miR-362-3p 和 Janus 激酶-2(Jak2)的水平。通过双荧光素酶报告基因实验、RNA 下拉实验和 RNA 免疫沉淀(RIP)实验验证 miR-362-3p 与 SNHG1 或 Jak2 的相互作用。此外,通过 Western blot 实验检测凋亡和自噬相关蛋白的水平。本研究中,LPS 抑制 PC12 细胞活力,并诱导其凋亡和自噬。SNHG1 敲低可影响细胞活力,并抑制 LPS 处理的 PC12 细胞的凋亡和自噬。此外,miR-362-3p 是 SNHG1 的靶标,miR-362-3p 靶向 Jak2 并负调控 Jak2/stat3 通路。我们的数据还表明,SNHG1 耗竭通过激活 Jak2/stat3 通路来影响 LPS 处理的 PC12 细胞中的细胞活力,并限制细胞凋亡和自噬,从而抑制 Jak2/stat3 通路。总之,SNHG1 通过海绵吸附 miR-362-3p 来调节 Jak2/stat3 通路,从而影响 LPS 处理的 PC12 细胞中的细胞活力、凋亡和自噬。