Antiangiogenesis effect of timosaponin AIII on HUVECs in vitro and zebrafish embryos in vivo

Authors: Zhong-yan Zhou1,2, Wai-rong Zhao1,3, Ying Xiao1, Xiang-ming Zhou1, Chen Huang2, Wen-ting Shi1, Jing Zhang1, Qing Ye1, Xin-lin Chen1, Jing-yi Tang1,3
1 Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
2 State Key Laboratory of Quality Research in Chinese Medicine and Institute of Chinese Medical Sciences, University of Macau, Macao, China
3 Cardiac rehabilitation Center of Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
Correspondence to: Xin-lin Chen:, Jing-yi Tang:,
DOI: 10.1038/s41401-019-0291-z
Received: 25 February 2019
Accepted: 17 July 2019
Advance online: 12 September 2019


Timosaponin AIII (Timo AIII) is a natural steroidal saponin isolated from the traditional Chinese herb Anemarrhena asphodeloides Bge with proved effectiveness in the treatment of numerous cancers. However, whether Timo AIII suppresses tumor angiogenesis remains unclear. In the present study, we investigated the antiangiogenesis effects of Timo AIII and the underlying mechanisms in human umbilical vein endothelial cells (HUVECs) in vitro and zebrafish embryos in vivo. We showed that treatment with Timo AIII (0.5–2 µM) partially disrupted the intersegmental vessels (ISVs) and subintestinal vessels (SIVs) growth in transgenic zebrafish Tg(fli-1a: EGFP)y1. Timo AIII (0.5–4 µM) dose-dependently inhibited VEGF-induced proliferation, migration, invasion, and tube formation of HUVECs, but these inhibitory effects were not due to its cytotoxicity. We further demonstrated that Timo AIII treatment significantly suppressed the expression of VEGF receptor (VEGFR) and the phosphorylation of Akt, MEK1/2, and ERK1/2 in HUVECs. Timo AIII treatment also significantly inhibited VEGF-triggered phosphorylation of VEGFR2, Akt, and ERK1/2 in HUVECs. Moreover, we conducted RNA-Seq and analyzed the transcriptome changes in both HUVECs and zebrafish embryos following Timo AIII treatment. The coexpression network analysis results showed that various biological processes and signaling pathways were enriched including angiogenesis, cell motility, cell adhesion, protein serine/threonine kinase activity, transmembrane signaling receptor activity, growth factor activity, etc., which was consistent with the antiangiogenesis effects of Timo AIII in HUVECs and zebrafish embryos. We conclude that the antiangiogenesis effect of Timo AIII is mediated through VEGF/PI3K/Akt/MAPK signaling cascade; Timo AIII potentially exerts antiangiogenesis effect in cancer treatment.
Keywords: Timosaponin AIII; Traditional Chinese herbal; Neovascularization; Neoplasm; VEGF/PI3K/Akt/MAPK; Transcriptome; Zebrafish; HUVECs; SU5416

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