589 SENP1 Knockdown Inhibits Hepatocyte Growth Factor Induced Migration and Epithelial Mesenchymal Transition of Hepatocellular Carcinoma Molecular Therapy Volume 23, Supplement 1, May 2015 Copyright[.]
Gene Inhibition ADSC were isolated from subcutaneous fat tissue of healthy young donors (n=4) and cultured up to 2-3 passages ADSC phenotype characterized by flow cytometry was CD90+/CD73+/CD105+/ CD45-/CD31- for all samples and these cells were capable of adipogenic and osteogenic differentiation We analyzed miR profile in ADSC using Illumina microarrays and found that miR-92a was one of the highly expressed miRs in all ADSC samples ADSC transfection by pre-miR-92a or anti-miR-92a led to the coordinated changes of miR-92a well-known target genes expression (integrin alpha 5, ITGA5, and mitogen activated protein kinase, MEK4) ADSC with over-expression of miR-92a completely lost the ability to stimulate capillary-like tube formation by endothelial cells (HUVEC) in vitro compared to ADSC transfected by the control gene However, transfection of ADSC by anti-miR-92a did not significantly increase their stimulating effect in tube formation assay As paracrine mechanisms are considered to be the most important for realizing of MSC angiogenic potential, we analyzed gene expression (qRT-PCR) and secretion (Bio-Plex assay and ELISA) of some key angiogenic factors in ADSC We found that mRNA level of vascular endothelial growth factor (VEGF), angiogenin and leptin was slightly increased in ADSC transfected by anti-miR-92a and this tendency was also observed for VEGF secretion When miR-92a was over-expressed in ADSC secretion of hepatocyte growth factor (HGF) and angiopoetin-1 by these cells significantly decreased compared to the control cells We conclude that miR-92a might play an important role in the regulation of ADSC ability to stimulate angiogenesis and key angiogenic factors such as VEGF, HGF and angiopoetin-1 are involved in this process The underlying mechanisms of miR92a-mediated direct or indirect effects in ADSC need to be further investigated Anyway, miR-92a could be considered as a promising target for the modulation of ADSC angiogenic potential 588 Effective Prevention of Liver Fibrosis by Liver-Targeted Hydrodynamic Gene Delivery of Matrix Metalloproteinase-13 in Rat Liver Fibrosis Model Hiroyuki Abe,1 Kenya Kamimura,1 Yuji Kobayashi,1 Masato Ohtsuka,2 Hiromi Miura,2 Riuko Ohashi,3 Takeshi Yokoo,1 Tsutomu Kanefuji,1 Takeshi Suda,1 Masanori Tsuchida,4 Yutaka Aoyagi,1 Guisheng Zhang,5 Dexi Liu,5 Shuji Terai.1 Division of Gastroenterology and Hepatology, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan; 2Department of Molecular Life Science, Division of Basic Molecular Science and Molecular Medicine, School of Medicine, Tokai University, Isehara, Kanagawa, Japan; 3Department of Pathology, Niigata University Medical and Dental Hospital, Niigata, Japan; 4Division of Thoracic and Cardiovascular Surgery, Graduate School of Medical and Dental Sciences, Niigata University, Niigata, Japan; 5Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, GA Liver fibrosis is final stage of chronic liver diseases which may cause liver failure and liver cancer While various diagnostic methods, including serum marker have been established, no standard therapy has developed The objective of this study is to assess the approach of overexpressing matrix metalloproteinase-13 gene (MMP13) in rat liver to block liver fibrosis The rat liver fibrosis model was developed by the bile duct ligation (BDL) And the liver-targeted hydrodynamic gene delivery procedure involves catheter insertion via the inferior vena cava (IVC) to the junction of the IVC and the hepatic veins followed by the hydrodynamic injection of MMP13 expression vector, containing CAG promoter-MMP13-IRES-tdTomato-polyA cassette with temporal blood flow occlusions at the supra- and infrahepatic IVC Three groups (n=5 in each group) of animals, BDL S234 alone (non-treated), BDL simultaneously followed by hydrodynamic delivery of pBGI-MMP13 (treated group), and hydrodynamic delivery of pBGI-MMP13 (control) were analyzed for: 1) MMP13 expression in serum and cells in the liver by ELISA, western blotting, immunohistochemical staining, and fluorescent detection; and 2) therapeutic effect on liver fibrosis by analysis of serum fibrotic markers of hyaluronic acid and collagen type IV, and histological analysis including silver staining Serum concentration of MMP13 in pBGI-MMP13 treated group reached 71.7±15.1 pg/ml (p