LncRNA SNHG1 protects SH-SY5Y cells from hypoxic injury through miR-140-5p/Bcl-XL axis
LncRNA SNHG1 protects SH-SY5Y cells from hypoxic injury through miR-140-5p/Bcl-XL axis
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DOI：
10.1080/00207454.2020.1744594
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发表时间：
2020-04-15
影响因子：
2.2
通讯作者：
Pang, Li
中科院分区：
文献类型：
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作者：
Wang, Wei;Lou, Xiao-Qian;Pang, Li
背景：缺氧性脑损伤是导致神经发育障碍和心血管功能障碍的主要原因之一。LncRNA SNHG1在缺氧性损伤中起关键作用，但其可能的作用机制尚不清楚。方法：采用定量逆转录聚合酶链式反应(qRT-PCR)检测小核仁RNA宿主基因1(SNHG1)和miR-140-5p的表达水平。免疫印迹法检测Bclxl和细胞凋亡相关蛋白的表达。采用双荧光素酶报告基因分析方法，检测lncRNA SNHG1与miR-140-5p、miR-140-5p与Bclxl之间的靶向关系。流式细胞仪Annexin V/PI染色检测细胞凋亡。用四甲基偶氮唑盐比色法检测细胞存活率。结果：氧糖剥夺(OGD)可抑制SNHG1和Bclxl的表达，增强miR-140-5p的表达。转染pcDNA3.1-SNHG1或miR-140-5p抑制剂后，OGD对SH-SY5Y细胞活力的抑制作用和对细胞凋亡的促进作用被部分取消。此外，荧光素酶报告实验表明，lncRNA SNHG1直接与miR-140-5p结合，miR-140-5p直接靶向Bcl-xL。在SH-SY5Y细胞中，转染miR-140-5p模拟物或sh-BclxL后，SNHG1过表达对OGD诱导的细胞凋亡的保护作用减弱。结论：lncRNA SNHG1介导的miR-140-5p抑制可调节Bclxl的表达，从而减少细胞凋亡，恢复SH-SY5Y细胞的活力。本研究结果为探讨SNHG1介导的信号通路在缺氧性脑损伤中的作用机制提供了新的思路。
Background: Hypoxic brain injury is one of the major causes of neurodevelopmental impairment and cardiovascular disability. LncRNA SNHG1 works as a critical factor in hypoxic induced injury, however, the potential mechanism is still not known well. Methods: The expression level of small nucleolar RNA host gene 1 (SNHG1) and miR-140-5p was detected by qRT-PCR. The western blot assay was performed to measure the level of Bcl-XL and apoptosis-related proteins. The target relationship between lncRNA SNHG1 and miR-140-5p, as well as miR-140-5p and Bcl-XL was detected by dual luciferase reporter gene assay. Cell apoptosis was assessed using Annexin V/PI staining by flow cytometry. Cell viability was analyzed by MTT assay. Results: Oxygen glucose deprivation (OGD) treatment inhibited SNHG1 and Bcl-XL expression and enhanced miR-140-5p expression. OGD treatment-induced cell viability inhibition, cell apoptosis promotion were partially abrogated when SH-SY5Y cells were transfected with pcDNA3.1-SNHG1 or miR-140-5p inhibitor. Moreover, luciferase reporter assay revealed that lncRNA SNHG1 bound directly to miR-140-5p, and miR-140-5p directly targeted Bcl-XL. The protective effect of SNHG1 overexpressing on cell apoptosis induced by OGD was attenuated after transfected with miR-140-5p mimic or sh-Bcl-XL in SH-SY5Y cells. Conclusion: LncRNA SNHG1-modulated miR-140-5p inhibition regulates Bcl-XL expression, thereby reducing cell apoptosis and recovering cell viability of SH-SY5Y cells. The results in this study provide novel insight into the mechanism of SNHG1 mediated signaling pathway during hypoxic brain injury.