Original article WNT5A/JNK signaling regulates pancreatic cancer cells migration by Phosphorylating Paxillin Wei Wei a , Hongyue Li a , Na Li a, b , Huihui Sun a , Qiang Li c , Xiaohong Shen a, b, * a School of Medicine, Nankai University, Tianjin 300071, China b Tianjin Key Laboratory of Tumor Microenvironment and Neurovascular Regulation, Tianjin 300071, China c Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China article info Article history: Received 11 January 2013 Received in revised form 29 May 2013 Accepted 29 May 2013 Keywords: WNT5A/JNK signaling Paxillin Pancreatic cancer cell Migration abstract Background: Expression of WNT5A associated with aggressive tumor biology and poor clinical outcome of various types of cancer. However its function in the metastasis property of pancreatic cells still needs to be elucidated. Methods: We detected the expressions of WNT5A, JNK1/p-JNK1 and Paxillin/p-Paxillin in cancer and the para-carcinoma tissues of pancreatic cancer. To understand how WNT5A/JNK signaling affects pancreatic cancer cell migration through the phosphorylation of cellular substrates of Paxillin, In vitro, we knocked down the WNT5A in PANC1, Capan-2 and HT1080 cell lines, and then tested the expression of JNK1. We detected the proteins of phosphorylation of Paxillin after JNK1 was inhibited and then the cells migration assay was evaluated. Moreover, JNK1 functionally phosphorylates serine178 on paxillin in vitro was detected .At last we subsequently observed whether WNT5A/JNK signaling modulates some molecule expressions relevant to focal adhesion (FA) formation and mesenchymal transition (EMT) and cell cycle. Results: WNT5A, p-JNK1 and p-Paxillin were highly expressed in early stage of tumor tissues. In vitro, WNT5A/JNK signaling promotes cell migration in pancreatic cancer by phosphorylating serine178 on Paxillin, an FA adaptor, which means WNT5A may regulate FA’s function.WNT5A up-regulates the molecule’s expressions relevant to cell adhesion through the phosphorylation of JNK1, including MMP1, MMP2, ICAM and CD44. In addition, WNT5A/JNK signaling promoted the mRNA expressions of vimentin, but decreased in E-Cadherin expression, which suggested its regulatory effects on the EMT processes. WNT5A/JNK signaling didn’t modulate cell proliferation. Conclusion: WNT5A/JNK signaling initiate cell migration of pancreatic cancer through activation of Paxillin, which suggested WNT5A has the potency of being an effective therapeutic target for the metastasis of pancreatic cancer. Copyright Ó 2013, IAP and EPC. Published by Elsevier India, a division of Reed Elsevier India Pvt. Ltd. All rights reserved. 1. Introduction Pancreatic cancer is the fourth most common cause of cancer death across the globe [1]. Because of the high metstatic property of tumor, pancreatic cancer patients always show high mortality rate. Thus better understanding of the molecular mechanism underlying metastasis is urgent for finding effective biomarkers of therapeutic targets in this tumor. Some proteins, such as KAI1, Matrix Metalloproteinases (MMPs), Vascular Endothelial Growth Factor (VEGF) and Tumor Growth Factor (TGF) [2e4], have been reported to be associated with the metastasis property of pancreatic cancer, but they are not specific for clinical treatment. Current researches revealed that tumor cells specifically secrete autocrine endogenous production into the extracellular matrix to stimulate their metastasis. Among them, the expression of WNT5A has been shown to be related with the cancer procession [5]. Particularly, WNT5A as a member of WNT family of secreted glycoproteins lacks transforming activity, thus contributes more to tumor progression than malignant initiation [6]. Also it has been reported that WNT5A can function either as a tumor sup- pressor or promoter depending on the type of cancer, which might constitute its determinant value for the progression of individual cancer [7]. Previous study has been found that WNT5A protein was associated with malignant metastasis, including pancreatic cancer [8e10]. However, the function of WNT5A in migration nature of * Corresponding author. School of Medicine, Nankai University, 94 Weijin Road, Tianjin 300071, China. Tel.: þ86 22 23501649; fax: þ86 22 23502554. E-mail addresses: shenxiaohong@nankai.edu.cn, shenxiaohong_2008@yahoo.cn (X. Shen). Contents lists available at SciVerse ScienceDirect Pancreatology journal homepage: www.elsevier.com/locate/pan 1424-3903/$ e see front matter Copyright Ó 2013, IAP and EPC. Published by Elsevier India, a division of Reed Elsevier India Pvt. Ltd. All rights reserved. http://dx.doi.org/10.1016/j.pan.2013.05.008 Pancreatology 13 (2013) 384e392