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Linc00299/miR-490-3p/AURKA轴调节动脉粥样硬化中的细胞生长和迁移。

Linc00299/miR-490-3p/AURKA axis regulates cell growth and migration in atherosclerosis.

作者信息

Liu Yong, Chen Yaqing, Tan Lili, Zhao Hongmei, Xiao Nuan

机构信息

Affiliated Hospital of Hebei University, No 212 Yuhua East Road, Baoding, 071000, Hebei, China.

出版信息

Heart Vessels. 2019 Aug;34(8):1370-1380. doi: 10.1007/s00380-019-01356-7. Epub 2019 Feb 8.

Abstract

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.

摘要

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。

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