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02 March 2020 : Laboratory Research  

Spalt-Like Protein 4 (SALL4) Promotes Angiogenesis by Activating Vascular Endothelial Growth Factor A (VEGFA) Signaling

Jinbo Sun1ABCDEF, Zhining Zhao2BCF, Wei Zhang1BF, Qisheng Tang1BF, Fan Yang1BF, Xiangnan Hu1CD, Chong Liu3CF, Bin Song1ADE*, Bo Zhang1ACE, He Wang1ADG

DOI: 10.12659/MSM.920851

Med Sci Monit 2020; 26:e920851

Abstract

BACKGROUND: Spalt-like protein 4 (SALL4) is a nuclear transcription factor central to early embryonic development, especially for regulating pluripotency of embryonic stem cells (ESCs) and sustaining ESCs self-renewal. Aberrant re-expression of SALL4 in adult tissues is involved in tumorigenesis and cancer progression. However, the role of SALL4 in angiogenesis remains elusive. Here, we determined the potential action of SALL4 on proliferation, migration, and tube formation of endothelial cells.

MATERIAL AND METHODS: HUVECs were infected with lentiviral particles expressing shRNA against SALL4. QRT-PCR and immunoblotting analysis were carried out to evaluate knockdown efficiency at mRNA and protein levels. Cell proliferation was measured by CCK-8 assay and flow cytometry was conducted to analyze cell cycle distribution. Wound-healing and Transwell migration assays were performed to evaluate cell motility. In addition, we determined the role of SALL4 on angiogenesis by tube formation assay, and Western blot analysis was used to assess the effect of SALL4 downregulation on VEGFA expression.

RESULTS: We found that SALL4 downregulation resulted in decreased proliferation. Cell cycle analysis revealed that SALL4 knockdown impeded cell cycle progression and induced cell cycle arrest at G1 phase. We also found that silencing of SALL4 decreased the capacity of wound healing and cell migration in HUVECs. Furthermore, tube formation assay showed that loss of SALL4 inhibited HUVECs angiogenesis. We also observed that SALL4 knockdown reduced the level of VEGFA in HUVECs.

CONCLUSIONS: In conclusion, these results support that by promoting proliferation, cell cycle progression, migration, and tube formation, SALL4 is involved in the process of angiogenesis through modulating VEGFA expression.

Keywords: Angiogenesis Inducing Agents, Cell Migration Assays, Vascular Endothelial Growth Factor A, Angiogenic Proteins, endothelial cells, G1 Phase, Gene Expression Regulation, Human Umbilical Vein Endothelial Cells, Neovascularization, Physiologic, Transcription Factors

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750