Involvement of miR-125b in human pancreatic ductal epithelial barrier affected by TRAF6

Abstract

Objective To investigate the effect of tumor necrosis factor receptor-associated factor 6 (TRAF6) on barrier and targeted binding of miR-125b to TRAF6 in human pancreatic ductal epithelial cell line HPDE6C7. Methods TRAF6 expression HPDE6C7 cells were up-regulated by recombinant lentivirus infection. The protein expression of nuclear factor κB(NF-κB) pathway and tight junction were examined by Western blot, the permeability of monolayer HPDE6C7 cells was measured with Transwell cab to evaluate the epithelial barrier function, and the targeted binding of miR-125b to TRAF6 was identified by dual-luciferase reporter assay. Results After up-regulation of TRAF6 expression by HPDE6C7 cells, the protein level of NF-κB p65, TGFβ-activated kinase 1(TAK1) and TAK1-binding 1 (TAB1) was increased, and the protein level of occludin and claudin-1 was decreased(P＜0.05), the permeability increased significantly (P＜0.05). Dual-luciferase reporter assay confirmed that TRAF6 was the target gene of miR-125b. Conclusions TRAF6 potentially play a role in affecting the pancreatic ductal epithelial barrier by regulating the NF-κB signaling pathway, which may be inhibited by miR-125b targeted binding to TRAF6.

Keywords

• acute pancreatitis|pancreatic ductal epithelial barrier|tumor necrosis factor receptor-associated factor 6 (traf6)|nuclear factor κb (nf-κb)|mir-125b