摘要： 目的 探讨长链非编码RNA（lncRNA）NEAT1在血管平滑肌细胞（VSMCs）增殖和凋亡中的作用及其信号传导机制。方法 应用实时定量PCR实验测定VSMCs中NEAT1和miR-490-3p的表达。CCK-8法分析NEAT1对VSMCs增殖的影响。流式细胞仪检测VSMCs的凋亡。通过免疫印迹实验检测VSMCs中hnRNPA1的表达水平。结果 在经氧化修饰的低密度脂蛋白处理的VSMCs中NEAT1表达以剂量依赖性和时间依赖性模式增加。NEAT1沉默抑制VSMCs的增殖并诱导了早期凋亡。si-NEAT1转染后miR-490-3p表达显著增高（P < 0.05），然而miR-490-3p的表达上调并不影响NEAT1表达（P > 0.05）。si-NEAT1转染VSMCs中hnRNPA1表达显著降低，同时在miR-490-3p过表达的VSMCs中hnRNPA1表达显著降低。结论 miR-490-3p是NEAT1的抑制靶标，过表达lncRNA NEAT1通过使miR-490-3p/hnRNPA1轴海绵化而促进VSMCs的增殖并且减少早期凋亡。

长链非编码RNA NEAT1对血管平滑肌细胞增殖和凋亡的影响及作用机制

孙骏, 肖亮, 宋健博

中国医科大学附属第四医院介入科, 沈阳 110032

收稿日期:2019-09-24出版日期:2020-07-30发布日期:2020-07-02
通讯作者:宋健博E-mail:jianbosong@cmu.edu.cn
作者简介:孙骏(1990-),男,医师,硕士.
基金资助:国家自然科学基金（81471763）


关键词: 长链非编码RNA, NEAT1, 血管平滑肌细胞, 增殖, 凋亡
Abstract: Objective To investigate the role of long non-coding RNA (lncRNA) NEAT1 on proliferation and apoptosis in vascular smooth muscle cells (VSMCs) and the signaling mechanism involved. Methods Quantitative real-time PCR (qRT-PCR) analysis was performed to measure the expression levels of lncRNA NEAT1 and miR-490-3p in VSMCs. The effect of lncRNA NEAT1 on the proliferation of VSMCs was analyzed using the CCK-8 assay. Flow cytometry analysis was performed to detect apoptosis in VSMCs. The expression of hnRNPA1 in VSMCs was assessed using Western blotting analysis. Results The expression levels of lncRNA NEAT1 were found to be increased in the ox-LDL induced VSMCs in a dose-and time-dependent manner. The knockdown of lncRNA NEAT1 led to the inhibition of proliferation and induction of early apoptosis in VSMCs. The expression of miR-490-3p was found to be significantly increased after si-NEAT1 transfection (P < 0.05);however,the up-regulation of miR-490-3p did not affect the expression of lncRNA NEAT1 (P > 0.05). Moreover,the expression levels of hnRNPA1 in VSMCs transfected with si-NEAT1 were found to be significantly reduced. The expression of hnRNPA1 in VSMCs that was induced by miR-490-3p was found to be significantly reduced after si-NEAT1 transfection. Conclusion miR-490-3p is an inhibitory target of NEAT1. The overexpression of lncRNA NEAT1 promotes the proliferation,and inhibits early apoptosis,in VSMCs by blocking the miR-490-3p/hnRNPA1 axis.
Key words: long non-coding RNA, NEAT1, vascular smooth muscle cell, proliferation, apoptosis
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