title: Knockdown of Decoy Receptor 3 Impairs Growth and Invasiveness of Hepatocellular Carcinoma Cell Line of HepG2.
abstract: BACKGROUND
  Decoy receptor 3 (DcR3) binds to Fas ligand (FasL) and inhibits FasL-induced apoptosis. The receptor is overexpressed in hepatocellular carcinoma (HCC), and it is associated with the growth and metastatic spread of tumors. DcR3 holds promises as a new target for the treatment of HCC, but little is known regarding the molecular mechanisms underlying the oncogenic properties of DcR3. The present work, therefore, examined the role of DcR3 in regulating the growth and invasive property of liver cancer cell HepG2.
  
  
  METHODS
  HepG2 cells were stably transfected with lentivirus-based short hairpin RNA vector targeting DcR3. After the knockdown of DcR3 was confirmed, cell proliferation, clone formation, ability of migrating across transwell membrane, and wound healing were assessed in vitro. Matrix metalloproteinase-9 (MMP 9) and vascular epithelial growth factor (VEGF)-C and D expressions of the DcR3 knockdown were also studied. Comparisons between multiple groups were done using one-way analysis of variance (ANOVA), while pairwise comparisons were performed using Student's t test. P< 0.05 was regarded statistically significant.
  
  
  RESULTS
  DcR3 was overexpressed in HepG2 compared to other HCC cell lines and normal hepatocyte Lo-2. Stable knockdown of DcR3 slowed down the growth of HepG2 (P < 0.05) and reduced the number of clones formed by 50% compared to those without DcR3 knockdown (P < 0.05). The knockdown also reduced the migration of HepG2 across transwell matrix membrane by five folds compared to the control (P < 0.05) and suppressed the closure of scratch wound (P < 0.05). In addition, the messenger RNA levels of MMP 9, VEGF-C, and VEGF-D were significantly suppressed by DcR3 knockdown by 90% when compared with the mock control (P < 0.05).
  
  
  CONCLUSIONS
  Loss of DcR3 impaired the growth and invasive property of HCC cell line of HepG2. Targeting DcR3 may be a potential therapeutic approach for the treatment of HCC.
authors: Department of General Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
authors: Xiao-Na
authors: XN
authors: Zhou
authors: Department of General Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
authors: Guang-Ming
authors: GM
authors: Li
authors: Department of General Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
authors: Ying-Chen
authors: YC
authors: Xu
authors: Department of General Surgery, Beijing Er Long Lu Hospital, Beijing 100032, China.
authors: Tuan-Jie
authors: TJ
authors: Zhao
authors: Department of General Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
authors: Ji-Xiang
authors: JX
authors: Wu
mesh_terms: D000704:Analysis of Variance; D002465:Cell Movement / Q000235:genetics / Q000502:physiology; D049109:Cell Proliferation / Q000235:genetics / Q000502:physiology; D056945:Hep G2 Cells; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D053323:Receptors, Tumor Necrosis Factor, Member 6b / Q000235:genetics / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D053323:Receptors, Tumor Necrosis Factor, Member 6b; D042461:Vascular Endothelial Growth Factor A; D020780:Matrix Metalloproteinase 9