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Notch activation suppresses endothelial cell migration and sprouting via miR-223-3p targeting Fbxw7

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Abstract

Angiogenesis involves temporo-spatially coordinated endothelial cell (EC) proliferation, differentiation, migration, and sprouting. Notch signaling is essential in regulating EC behaviors during angiogenesis, but its downstream mechanisms remain incompletely defined. In the current study, we show that miR-223-3p is a downstream molecule of Notch signaling and mediates the role of Notch signaling in regulating EC migration and sprouting. In human umbilical vein endothelial cells (HUVECs), Notch activation by immobilized Dll4, a Notch ligand, upregulated miR-223-3p, and Notch activation–mediated miR-223-3p upregulation could be blocked by a γ-secretase inhibitor (DAPT). miR-223-3p overexpression apparently repressed HUVEC migration, leading to attenuated lumen formation and sprouting capacities. Transcriptome comparison and subsequent qRT-PCR validation further indicated that miR-223-3p downregulated the expression of multiple genes involved in EC migration, axon guidance, extracellular matrix remodeling, and angiogenesis. In addition, miR-223-3p antagonist transfection abolished Notch-mediated repression of EC migration and sprouting. By quantitative reverse transcription–polymerase chain reaction (qRT-PCR), western blotting, and reporter assay analysis, we confirmed that miR-223-3p directly targeted F-box and WD repeat domain–containing 7 (Fbxw7). Meanwhile, Fbxw7 overexpression could efficiently rescue the impaired migration capacity of ECs under miR-223-3p overexpression. In summary, these results identify that Notch activation–induced miR-223-3p suppresses EC migration and sprouting via Fbxw7.

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Funding

This study was supported by the National Natural Science Foundation of China (31671523, 31730041, 82003110) and Natural Science Foundation of Shaanxi Province (2020JQ-441).

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Authors and Affiliations

  1. Faculty of Life Sciences, Northwest University, Xi’an, 710069, China

    Ruonan Wang, Xingxing Feng & Bo Che

  2. State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Chang-Le Xi Street #169, Xi’an, 710032, China

    Ruonan Wang, Ziyan Yang, Liang Liang, Xingxing Feng, Bo Che, Xiaoyan Zhang, Xianchun Yan & Hua Han

  3. Department of Cardiovascular Surgery, Shenzhen People’s Hospital, the Second Clinical Medical School, Jinan University & the First Affiliated Hospital, Southern University of Science and Technology, Shenzhen, 518020, China

    Qijun Zheng

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  1. Ruonan Wang
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Contributions

R.N.W. and Z.Y.Y. performed experiments and collected data. L.L., X.X.F., B.C., and X.Y.Z. assisted with experiments and data collection. H.H., X.C.Y., and Q.J.Z. designed the experiments and prepared the manuscript.

Corresponding authors

Correspondence to Qijun Zheng, Xianchun Yan or Hua Han.

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The use of human samples was approved by the Ethics Committee of Xijing Hospital, Fourth Military Medical University. All human participants signed informed consent for the use of their umbilical cord biopsies. The animal experiments were permitted by the Animal Experiment Administration Committee of the Fourth Military Medical University.

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The authors declare no competing interests.

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Ruonan Wang and Ziyan Yang are contributed equally to this study

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Wang, R., Yang, Z., Liang, L. et al. Notch activation suppresses endothelial cell migration and sprouting via miR-223-3p targeting Fbxw7. In Vitro Cell.Dev.Biol.-Animal 58, 124–135 (2022). https://doi.org/10.1007/s11626-022-00649-y

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