Study on the mechanism of miRNA-129-5p in epithelial-mesenchymal transition of biliary atresia mouse / 重庆医学

Objective To investigate the mechanism of miRNA-129-5p in epithelial-mesenchymal transition (EMT)of biliary atresia.Methods Constructed bile duct epithelial EMT cell model (the experimental group) induced by TGF-β1,detected the expressions of EMT related markers and miRNA-129-5p.While miRNA-129-5p precursor was transfected,the expressions of EMT related markers and extracellular matrix were contrasted between the original and the renovated biliary epithelial cells.Results In the experimental group,extrahepatic bile duct showedEMT,the expression of miRNA-129-5p was decreased (P＜0.05),overexpression of miRNA-129-5p could inhibit the progression of EMT (P＜0.05).Conclusion miRNA-129-5p may relate to EMT by regulating the expression of TGF-β1.

miRNA-129-5p regulates the targets of the VCP gene expression in human osteosarcoma cells / 医学研究生学报

Objective High expression of the valosin-containing protein ( VCP) gene can enhance the metastasis of osteosar-coma via the AKT/PI3K/NF-KappaB/MMP-9 signaling pathway, but the molecular mechanisms underlying the up-regulation of VCP in osteosarcoma cells remains unknown .This study aimed to determine whether miRNA-129-5p can regulate the VCP expression and its targets in human osteosarcoma cells . Methods The microRNA target-predicting software TargetScanhuman 6.2 ( http://www.tar-getscan.org/) was used to predict the possible targets of miRNA-129-5p on the VCP gene.Then, two recombinant gene report vectors containing the wild VCP gene 3′UTR ( psi-VCP vector ) and mutant VCP gene 3′UTR ( psi-VCPmut vector ) were constructed , se-quenced, and identified.The human osteosarcoma U2-OS cells were co-transfected with miRNA-129-5p mimic and psi-VCP vector or psi-VCPmut vector, respectively.A non-specificity mimic transfection served as negative control , and the luciferase activity was detec-ted in each group. Results The software prediction showed only one conserved function site of miRNA-129-5p on the VCP gene 3′UTR163-169 bp.Luciferase activity was significantly lower in the psi-VCP vector +miRNA-129-5p transfection group (15.529 ± 1.902) than in the VCP control group (21.781 ±0.854), VCP mutation experimental group (19.978 ±1.377), and VCP mutation control group (21.952 ±1.516) (P<0.05), with no remarkable difference between the VCP mutation control and VCP control groups (P=0.276). Conclusion miRNA-129-5p can probably regulate the targets of the VCP gene in human osteosarcoma U 2-OS cells.