Effect of miR-144-5p on the proliferation, migration, invasion and apoptosis of human umbilical vein endothelial cells by targeting RICTOR and its related mechanisms
Effect of miR-144-5p on the proliferation, migration, invasion and apoptosis of human umbilical vein endothelial cells by targeting RICTOR and its related mechanisms
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DOI：
10.3892/etm.2019.8369
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发表时间：
2020-03-01
影响因子：
2.7
通讯作者：
Zhao, Heng
中科院分区：
文献类型：
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作者：
Fu, Wei;Liu, Zidong;Zhao, Heng
本研究旨在观察microRNA（miR）-144-5p对人脐静脉内皮细胞（HUVECs）的影响，探讨miR-144- 5 p在动脉粥样硬化中的作用。使用miR-144 - 5 p模拟物在HUVEC中上调miR-144 - 5 p表达。逆转录-定量PCR（RT-qPCR）检测HUVECs中miR-144- 5 p的相对表达水平。MTT法检测细胞增殖情况。通过流式细胞术测定细胞凋亡。通过伤口愈合试验检测细胞迁移能力。通过transwell测定确定细胞侵袭。Western blot检测磷酸化（p）-PI 3 K、p-Akt和内皮型一氧化氮合酶（eNOS）蛋白水平。通过TargetScan预测miR-144- 5 p与雷帕霉素不敏感的mTOR伴体（RICTOR）mRNA 3 '-非翻译区的结合位点，并通过双荧光素酶报告基因分析证实。目前的研究表明，miR-144- 5 p模拟物显著抑制HUVECs的细胞增殖并诱导凋亡。此外，miR-144- 5 p模拟物可抑制HUVECs的迁移和侵袭。进一步分析表明RICTOR是miR-144- 5 p的直接靶基因。此外，miR-144- 5 p上调降低了p-PI 3 K、p-Akt和eNOS的蛋白水平。总之，miR-144- 5 p通过改变RICTOR的表达来影响PI 3 K-Akt-eNOS信号通路，从而调节HUVEC的增殖、迁移、侵袭和凋亡。这些结果表明miR-144- 5 p可能是预防和治疗动脉粥样硬化的潜在靶点。
The purpose of the present study was to investigate the effect of microRNA (miR)-144-5p on human umbilical vein endothelial cells (HUVECs) to explore the role of miR-144-5p in atherosclerosis. miR-144-5p expression was upregulated in HUVECs using miR-144-5p mimics. The relative expression level of miR-144-5p in HUVECs was detected using reverse transcription-quantitative PCR (RT-qPCR). Cell proliferation was detected by performing an MTT assay. Apoptosis was determined via flow cytometry. Cell migration ability was detected by a wound-healing assay. Cell invasion was determined by a transwell assay. The protein levels of phosphorylated (p)-PI3K, p-Akt and endothelial nitric oxide synthase (eNOS) were detected using western blot analysis. The binding sites between miR-144-5p and 3 '-untranslated region of rapamycin-insensitive companion of mTOR (RICTOR) mRNA were predicted by TargetScan and confirmed by a dual luciferase reporter assay. The present study showed that miR-144-5p mimics significantly inhibited cell proliferation and induced apoptosis in HUVECs. In addition, miR-144-5p mimics could suppress migration and invasion of HUVECs. Further analysis identified that RICTOR was a direct target gene of miR-144-5p. Moreover, miR-144-5p upregulation decreased the protein level of p-PI3K, p-Akt and eNOS. In conclusion, miR-144-5p regulated HUVEC proliferation, migration, invasion, and apoptosis through affecting the PI3K-Akt-eNOS signaling pathway by altering the expression of RICTOR. These results indicated that miR-144-5p may be a potential target for the prevention and treatment of atherosclerosis.