Liu Yong, Chen Yaqing, Tan Lili, Zhao Hongmei, Xiao Nuan
Affiliated Hospital of Hebei University, No 212 Yuhua East Road, Baoding, 071000, Hebei, China.
Heart Vessels. 2019 Aug;34(8):1370-1380. doi: 10.1007/s00380-019-01356-7. Epub 2019 Feb 8.
Long non-coding RNA (lncRNA) plays a crucial role in regulating various cellular processes in atherosclerosis. The present study identified the regulation of Linc00299, via miR-490-3p targeting Aurora kinase A (AURKA), on migration and proliferation of endothelial cells and vascular smooth muscle cells (VSMCs) during atherosclerosis. The expression of RNAs was assessed by real-time PCR. The proliferation, apoptosis and migration were detected using MTT assay, Annexin V/PI staining and Transwell system, respectively. Bindings of Linc00299/miR-490-3p and subsequent miR-490-3p/AURKA were verified by luciferase and biotin pull-down assays. The protein expression of AURKA was detected by Western blotting. Expressions of Linc00299 and miR-490-3p were upregulated and downregulated in atherosclerosis patients, respectively. Both Linc00299 knockdown and miR-490-3p overexpression suppressed cell proliferation, increased apoptosis and inhibited migration of VSMCs and HUVECs. Linc00299 directly bound to miR-490-3p which targeted AURKA. The regulation of Linc00299 on expression of AURKA and proliferation and migration of VSMCs were dependent on miR-490-3p. Atherosclerosis-increased Linc00299 acts as a sponge of miR-490-3p to upregulate AURKA, and as a result increases proliferation and migration in VSMCs and HUVECs. Our study reveals an important effect of Linc00299/miR-490-3p/AURKA axis on regulating cell proliferation and migration in atherosclerosis.
长链非编码RNA(lncRNA)在动脉粥样硬化中调节各种细胞过程中起着关键作用。本研究确定了通过miR-490-3p靶向极光激酶A(AURKA)的Linc00299对动脉粥样硬化过程中内皮细胞和血管平滑肌细胞(VSMC)迁移和增殖的调节作用。通过实时PCR评估RNA的表达。分别使用MTT法、Annexin V/PI染色和Transwell系统检测增殖、凋亡和迁移。通过荧光素酶和生物素下拉试验验证Linc00299/miR-490-3p以及随后的miR-490-3p/AURKA的结合。通过蛋白质印迹法检测AURKA的蛋白表达。在动脉粥样硬化患者中,Linc00299和miR-490-3p的表达分别上调和下调。Linc00299敲低和miR-490-3p过表达均抑制了VSMC和HUVEC的细胞增殖,增加了细胞凋亡并抑制了其迁移。Linc00299直接与靶向AURKA的miR-490-3p结合。Linc00299对AURKA表达以及VSMC增殖和迁移的调节依赖于miR-490-3p。动脉粥样硬化中增加的Linc00299充当miR-490-3p的海绵以上调AURKA,结果增加了VSMC和HUVEC中的增殖和迁移。我们的研究揭示了Linc00299/miR-490-3p/AURKA轴在调节动脉粥样硬化中细胞增殖和迁移方面的重要作用。