Abstract:

Objective To investigate the role and possible mechanism of miR-590 in epithelial-mesenchymal transition （EMT） of cholangiocarcinoma cells. Methods  Fluorescence quantitative PCR （QPCR） was used to detect the expression of miR-590 in human normal intrahepatic bile duct epithelial cell line HIBEC， human cholangiocarcinoma cell line HUCCT1， HCCC-9810 and RBE. Dual luciferase reporter assay was used to evaluate the regulatory effect of miR-590 on SIP1. MiR-590 mimics （transfected group） and nonsense nucleic aicd sequence （NC group） were transfected into HUCCT1 cells， and Western blotting was used to detect the expression of EMT related protein in two groups. SIP1 siRNA was transfected into HUCCT1 cells and Western blotting was used to detect EMT related protein expression after interfering SIP1. Results The expression levels of miR-590 in HUCCT1， HCCC-9810 and RBE cell lines were 0.37±0.084，0.31±0.071 and 0.53±0.089， which was significantly lower than that of normal intrahepatic bile duct epithelial cell line HIBEC （1.12±0.201）， and the difference was statistically significant （P＜0.05）. MiR-590 mimics significantly reduced luciferase expression in the psiCHECK2-SIP1-3’UTR group （0.37±0.041， P＜0.001），while not affecting the expression of the psiCHECK2-SIP1-3’UTR-MUT group （1.21±0.211， P＞0.05）. Overexpressed of miR-590 could down-regulated the mesenchymal proteins Vimentin， N-cadherin， ZEB1， ETS1， SNAIL1， TWIST1 and up-regulated the epithelial marker E-cadherin. SIP1 silencing could up-regulate the expression of E-cadherin protein and down-regulate the expression of Vimentin and N-cadherin protein. Conclusion MiR-590 blocked its translation by targeting SIP1-3’UTR and ultimately inhibited epithelial mesenchymal transition of cholangiocarcinoma cells.

Key words:

Cholangiocarcinoma；miR-590；SIP1；Epithelial-mesenchymal transition（EMT）  ')">"> Cholangiocarcinoma；miR-590；SIP1；Epithelial-mesenchymal transition（EMT）  ,  ,  ,