Esculetin Inhibits VEGF-Induced Angiogenesis BothIn VitroandIn Vivo

Abstract

Esculetin is known to inhibit tumor growth, but its effect in angiogenesis has not been studied. Here, we report the efficacy of esculetin on VEGF-induced angiogenesis. Esculetin treatment inhibited VEGF-induced proliferation and DNA synthesis of HUVECs with no cell toxicity. G1-phase cell-cycle arrest was associated with a decreased expression of cyclins and CDKs via the binding of p27KIP1. Esculetin down-regulated the MMP-2 expression in VEGF-stimulated HUVECs, which suppressed colony tube formation and migration. Esculetin reduced the phosphorylation of VEGFR-2 and the downstream signaling of VEGFR-2, including ERK1/2 and eNOS/Akt pathways. Esculetin suppressed microvessel outgrowth from an aortic ring ex vivo model treated with VEGF, and blocked the VEGF-induced formation of new blood vessels and hemoglobin content in an in vivo Matrigel plug model. Collectively, VEGF-stimulated responses in angiogenesis were inhibited in vitro and in vivo, providing a theoretical basis for effective use against anti-angiogenic therapies.

Phytochemicals Targeting VEGF and VEGF-Related Multifactors as Anticancer Therapy

“…Esculetin, a coumarin derivative, inhibits VEGF-induced proliferation and DNA synthesis, as well as downregulates MMP9 expression in HUVECs. Furthermore, esculetin reduces VEGFR2 phosphorylation and downregulates the ERK1/2 and eNOS/Akt signaling pathways, showing its potential role in inhibiting cancer progression [126].…”

Hydrangenol suppresses VEGF-stimulated angiogenesis by targeting p27KIP1-dependent G1-cell cycle arrest, VEGFR-2-mediated signaling, and MMP-2 expression

“…The cells were fixed and stained with propidium iodide (PI) and subjected to cell cycle analysis as reported previously (Park et al. 2016). The phase distribution of the cell cycle was measured using FACStar flow cytometer (BD Biosciences, San Jose, CA) equipped with the BD Cell Fit software.…”

“…To investigate protein effectors associated hydragenol activity, western blotting, immunoprecipitation, and immune complex kinase assays were performed as described previously (Park et al. 2016).…”

“…To exclude the possibility of proliferation-mediated migration, cells were pre-incubated with 5 μg/mL of mitomycin C (Sigma-Aldrich) for 2 h. Thereafter, hydrangenol-mediated cellular migration and invasion were assessed and gelatinase activity was measured via zymography, as reported previously (Park et al. 2016). Morphological changes in cells after triacanthine treatment were confirmed via microscopy images acquired using an inverted microscope at 40× magnification.…”

Topical administration of Esculetin as a potential therapy for experimental dry eye syndrome

“…Esculetin, which was extracted from Chinese herb drug 'Qinpi', was known for inducing apoptosis in human cancer cells, 4 attenuated the expression of pro-inflammatory mediators in the inflamed colon 5 and inhibiting VEGF-induced cell proliferation and DNA synthesis with no cell toxicity both in vitro and in vivo 6 in recent studies. Researches on Esculetin were systemic administration in mice or rats, while topical Esculetin in curing ocular diseases had not ever been reported before.…”