Crucial relationship of Neural Wiskott Aldrich syndrome protein(N-WASP) and Lysyl oxidase-like 2(LOXL2) in the promotion of pancreatic cancer metastasis

Abstract

Introduction: Pancreatic cancer has been widely known to have aggressive malignancy and poor prognosis. N-WASP is part of the WASP family that regulate actin polymerization by activating Arp2/3 complex. FAK(Focal adhesion kinase) promotes N-WASP and epithelial mesenchymal transition(EMT) in cancer metastasis. The overexpression of LOXL2 promotes distant metastasis in pancreatic cancer. However, there is insufficient evidence of the relationship between LOXL2 and N-WASP in pancreatic cancer. In this study, we aimed to confirm the expression of N-WASP and relation between LOXL2 and N-WASP in pancreatic cancer. Method: Between June 2002 and December 2012, 81 patients underwent radical curative resection for pancreatic cancer at Gangnam Severance Hospital. Pancreatic cancer cell lines MIA PaCa-2, PANC-1 were used for in vitro study. To evaluate the endogenous expression level of N-WASP, we purified the whole RNA and protein to perform the qPCR, RT-PCR and Western blot. And we confirmed the motility and invasiveness of MIA PaCa-2 and PANC-1. Result: High expression of LOXL2 and N-WASP was observed and correlated with the expression of mesenchymal markers (Snail, L1CAM, Vimentin). We generated stably LOXL2 silenced pancreatic cancer cell which exhibited significant changes of factors related to invasiveness and N-WASP expression. Importantly, silencing of N-WASP markedly decrease in motility and invasiveness of MIA PaCa-2 and PANC-1 as determined from wound healing assay. Conclusion: Our results demonstrated that N-WASP is a regulator of EMT for migration and invasion in pancreatic cancer. LOXL2 and N-WASP have a powerful relationship for activating the promotion of EMT in pancreatic cancer metastasis. These findings suggest that N-WASP can be a target for the determining of distant metastasis in pancreatic cancer.