Aeroallergen Der p 2 promotes motility of human non-small cell lung cancer cells via toll-like receptor-mediated up-regulation of urokinase-type plasminogen activator and integrin/focal adhesion kinase signaling

Abstract

// Chun-Hsiang Lin 1, * , Hui-Han Lin 1, 2, * , Cheng-Yi Kuo 3 , Shao-Hsuan Kao 1, 4 1 Institute of Biochemistry, Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan 2 Surgical Department Cardiovascular Division, China Medical University Hospital, Taichung, Taiwan 3 Department and Graduate Institute of Biology and Anatomy, National Defense Medical Center, Taipei, Taiwan 4 Clinical Laboratory, Chung Shan Medical University Hospital, Taichung, Taiwan * These authors contributed equally to this work Correspondence to: Shao-Hsuan Kao, email: kaosh@csmu.edu.tw Keywords: Der p 2, non-small cell lung cancer, integrin, urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor Received: August 12, 2016      Accepted: December 27, 2016      Published: January 05, 2017 ABSTRACT House dust mite (HDM) allergens are one of the major causes leading to respiratory hypersensitiveness and airway remodeling. Here we hypothesized that a major HDM allergen Der p 2 could increase cell motility and invasiveness of non-small cell lung cancer (NSCLC) cells. Our results showed that low dose (1 and 3 μg/mL) recombinant Der p 2 protein (DP2) enhanced the migration and invasiveness of human NSCLC cell A549, H1299 and CL1-5, but nonsignificantly altered their growth. Further investigation revealed that integrin αV level was increased and its downstream signaling including focal adhesion kinase (FAK) and paxillin were activated in A549 cells exposed to DP2. In parallel, DP2 also activated the FAK-associated signaling effectors such as Src, phosphatidyl inositol 3-kinase (PI3K), AKT, p38 mitogen-activated protein kinase (P38), extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK). Our findings also revealed that DP2 increased expression level of urokinase type plasminogen-activated kinase (uPA) and uPA receptor (uPAR), and subsequently enhanced the binding of uPAR to integrin αV. Moreover, the involvement of toll-like receptor 2/4 (TLR2/4)-triggered ERK1/2 activation in the increased expression of uPA and uPAR was also demonstrated. Collectively, these findings indicate that DP2 can enhance cell motility and invasiveness of NSCLC cells, attributing to TLR2/4-ERK1/2 activation, increased uPA and uPAR expression, enhanced binding of uPAR to integrin αV, and the consequent FAK signaling cascades. Thus, we suggest that DP2 may exacerbate NSCLC via promoting metastatic ability of carcinoma cell.