WMJ-S-001, a Novel Aliphatic Hydroxamate-Based Compound, Suppresses Lymphangiogenesis Through p38mapk-p53-survivin Signaling Cascade.

Shiu-Wen Huang,Ming-Jen Hsu,Wei-Chuan Chen,Hung-Yu Yang,Meng-Chieh Yu,Shih-Wei Wang,Ya-Fen Hsu,Wei-Jan Huang

doi:10.3389/fonc.2019.01188

Abstract

Background and purpose: Angiogenesis and lymphangiogenesis are major routes for metastatic spread of tumor cells. It thus represent the rational targets for therapeutic intervention of cancer. Recently, we showed that a novel aliphatic hydroxamate-based compound, WMJ-S-001, exhibits anti-angiogenic, anti-inflammatory and anti-tumor properties. However, whether WMJ-S-001 is capable of suppressing lymphangiogenesis remains unclear. We are thus interested in exploring WMJ-S-001's anti-lymphangiogenic mechanisms in lymphatic endothelial cell (LECs).Experimental approach: WMJ-S-001's effects on LEC proliferation, migration and invasion, as well as signaling molecules activation were analyzed by immunoblotting, flow-cytometry, MTT, BrdU, migration and invasion assays. We performed tube formation assay to examine WMJ-S-001's ex vivo anti-lymphangiogenic effects.Key results: WMJ-S-001 inhibited serum-induced cell proliferation, migration, invasion in murine LECs (SV-LECs). WMJ-S-001 reduced the mRNA and protein levels of survivin. Survivin siRNA significantly suppressed serum-induced SV-LEC invasion. WMJ-S-001 induced p53 phosphorylation and increased its reporter activities. In addition, WMJ-S-001 increased p53 binding to the promoter region of survivin, while Sp1 binding to the region was decreased. WMJ-S-001 induced p38 mitogen-activated protein kinase (p38MAPK) activation. p38MPAK signaling blockade significantly inhibited p53 phosphorylation and restored survivin reduction in WMJ-S-001-stimulated SV-LCEs. Furthermore, WMJ-S-001 induced survivin reduction and inhibited cell proliferation, invasion and tube formation of primary human LECs.Conclusions and Implications: These observations indicate that WMJ-S-001 may suppress lymphatic endothelial remodeling and reduce lymphangiogenesis through p38MAPK-p53-survivin signaling. It also suggests that WMJ-S-001 is a potential lead compound in developing novel agents for the treatment of lymphangiogenesis-associated diseases and cancer.

Highlights

Lymphangiogenesis, the formation of new lymphatic vessels, occurs primarily in many physiological processes such as embryonic development, tissue repair and resolution of inflammatory reactions
WMJ-S-001 Inhibits Lymphangiogenesis via Survivin Reduction p38 mitogen-activated protein kinase (p38MAPK)-p53-survivin signaling. It suggests that WMJ-S-001 is a potential lead compound in developing novel agents for the treatment of lymphangiogenesisassociated diseases and cancer
Antibodies against PARP, caspase 3 active form, survivin, ERK1/2 and ERK1/2 phosphorylated at Thr 202/Tyr 204, p38MAPK, p38MAPK phosphorylated at Thr180/Tyr182, p53 phosphorylated at Ser15 and p53 acetylated at Lys379 were from Cell Signaling (Danvers, MA, U.S.A.)

Summary

Introduction

Lymphangiogenesis, the formation of new lymphatic vessels, occurs primarily in many physiological processes such as embryonic development, tissue repair and resolution of inflammatory reactions. A variety of mechanisms such as seeding of body cavities, local tissue invasion, invasion into lymphatics and hematogenous spread are involved in metastatic tumor spread. Both blood and lymphatic systems contribute to tumor progression and metastasis, the dissemination of tumor cells via lymphatic vasculature is the most common route for most carcinomas [3, 4]. Angiogenesis and lymphangiogenesis are major routes for metastatic spread of tumor cells. It represent the rational targets for therapeutic intervention of cancer. We are interested in exploring WMJ-S-001’s anti-lymphangiogenic mechanisms in lymphatic endothelial cell (LECs). We performed tube formation assay to examine WMJ-S-001’s ex vivo anti-lymphangiogenic effects

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