WISP-1 promotes VEGF-C-dependent lymphangiogenesis by inhibiting miR-300 in human oral squamous cell carcinoma cells

// Ching-Chia Lin 1 , Po-Chun Chen 2, 3 , Ming-Yu Lein 2, 4 , Ching-Wen Tsao 2 , Chiu-Chen Huang 5 , Shih-Wei Wang 6 , Chih-Hsin Tang 2, 7, 8 , Kwong-Chung Tung 1 1 Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan 2 Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan 3 Department of Medical Research, Chung Shan Medical University Hospital, Chung Shan Medical University, Taichung, Taiwan 4 Division of Hematology and Oncology, Department of Internal Medicine, China Medical University Hospital, Taichung, Taiwan 5 Sing Wang Animal Hospital, Taichung, Taiwan 6 Department of Medicine, Mackay Medical College, New Taipei City, Taiwan 7 Department of Pharmacology, School of Medicine, China Medical University, Taichung, Taiwan 8 Department of Biotechnology, College of Health Science, Asia University, Taichung, Taiwan Correspondence to: Kwong-Chung Tung, e-mail: vettung@gmail.com Chih-Hsin Tang, e-mail: chtang@mail.cmu.edu.tw Keywords: WISP-1, OSCC, lymphangiogenesis, VEGF-C, miR-300 Received: August 12, 2015      Accepted: January 01, 2016      Published: January 25, 2016 ABSTRACT Oral squamous cell carcinoma (OSCC), which accounts for nearly 90% of head and neck cancers, is characterized by a poor prognosis and a low survival rate. Vascular endothelial growth factor-C (VEGF-C) has been implicated in lymphangiogenesis and is correlated with cancer metastasis. WNT1-inducible signaling pathway protein-1 (WISP)-1/CCN4 is an extracellular matrix-related protein that belongs to the CCN family and stimulates many biological functions. Our previous studies showed that WISP-1 plays an important role in OSCC migration and angiogenesis. However, the effect of WISP-1 on VEGF-C regulation and lymphangiogenesis in OSCC is poorly understood. Here, we showed a correlation between WISP-1 and VEGF-C in tissue specimens from patients with OSCC. To examine the lymphangiogenic effect of WISP-1, we used human lymphatic endothelial cells (LECs) to mimic lymphatic vessel formation. The results showed that conditioned media from WISP-1-treated OSCC cells promoted tube formation and cell migration in LECs. We also found that WISP-1-induced VEGF-C is mediated via the integrin αvβ3/integrin-linked kinase (ILK)/Akt signaling pathway. In addition, the expression of microRNA-300 (miR-300) was inhibited by WISP-1 via the integrin αvβ3/ILK/Akt cascade. Collectively, these results reveal the detailed mechanism by which WISP-1 promotes lymphangiogenesis via upregulation of VEGF-C expression in OSCC. Therefore, WISP-1 could serve as therapeutic target to prevent metastasis and lymphangiogenesis in OSCC.