title: The significance of Notch1 compared with Notch3 in high metastasis and poor overall survival in hepatocellular carcinoma.
abstract: BACKGROUND
  The prognosis for patients with hepatocellular carcinoma (HCC) is poor, and the mechanisms underlying the development of HCC remain unclear. Notch1 and Notch3 may be involved in malignant transformation, although their roles remain unknown.
  
  
  METHODS
  HCC tissues were stained with anti-Notch1 or -Notch3 antibody. The migration and invasion capacities of the cells were measured with transwell cell culture chambers. RT-PCR was used to measure the expression of Notch1 and Notch3 mRNA. Additionally, western blot analysis was used to assess the protein expression of Notch1, Notch3, CD44v6, E-cadherin, matrix metalloproteinase-2 (MMP-2), MMP-9, and urokinase-type plasminogen activator (uPA). RNA interference was used to down-regulate the expression of Notch1 and Notch3. Cell viability was assessed using MTT.
  
  
  RESULTS
  Based on immunohistochemistry, high Notch1 expression was correlated with tumor size, tumor grade, metastasis, venous invasion and AJCC TNM stage. High Notch3 expression was only strongly correlated with metastasis, venous invasion and satellite lesions. Kaplan-Meier curves demonstrated that patients with high Notch1 or Notch3 expression were at a significantly increased risk for shortened survival time. In vitro, the down-regulation of Notch1 decreased the migration and invasion capacities of HCC cells by regulating CD44v6, E-cadherin, MMP-2, MMP-9, and uPA via the COX-2 and ERK1/2 pathways. Down-regulation of Notch3 only decreased the invasion capacity of HCC cells by regulating MMP-2 and MMP-9 via the ERK1/2 pathway.
  
  
  CONCLUSIONS
  Based on the migration and invasion of HCC, we hypothesize that targeting Notch1 may be more useful than Notch3 for designing novel preventive and therapeutic strategies for HCC in the near future.
authors.affiliation: Department of Hepatobiliary Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shannxi, People's Republic of China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhou
authors.forename: Ning
authors.initials: N
authors.lastname: Zhang
authors.forename: Wenjie
authors.initials: W
authors.lastname: Song
authors.forename: Nan
authors.initials: N
authors.lastname: You
authors.forename: Qingjun
authors.initials: Q
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Desheng
authors.initials: D
authors.lastname: Wang
authors.forename: Kefeng
authors.initials: K
authors.lastname: Dou
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D017931:DNA Primers; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000502:physiology*; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016019:Survival Analysis
chemical_list: D017931:DNA Primers; C497761:NOTCH1 protein, human; C499374:NOTCH3 protein, human; D051881:Receptor, Notch1; D000071656:Receptor, Notch3; D051880:Receptors, Notch