title: Low-molecular weight chitosan/vascular endothelial growth factor short hairpin RNA for the treatment of hepatocellular carcinoma.
abstract: OBJECTIVE
  Vascular endothelial growth factor (VEGF) has been shown to be a key driving force for angiogenesis and tumor growth in hepatocellular carcinoma (HCC). As an emerging approach to block this angiogenic stimulator, the RNA interference (RNAi) technique has rapidly developed but is hindered for in vivo applications due to low cellular uptake and poor stability of small RNA. Based on low molecular weight chitosan (LMWC), a gene delivery system of short hairpin RNA (shRNA) directed against VEGF was constructed. The objective of this study was to investigate whether LMWC/shRNA nano-complexes can effectively inhibit VEGF expression in cancer cells and tumor tissues and suppress tumor growth in different HCC models.
  
  
  METHODS
  The transfection experiment and Real-time qPCR assay were used to evaluate the transfection efficiency and gene suppression activity of LMWC/shRNA complexes in Hepa 1-6 murine hepatocarcinoma cells. The therapeutic effect of LMWC/ VEGF shRNA was further tested in ectopic and orthotopic liver cancer models.
  
  
  RESULTS
  LMWC/VEGF shRNA complexes significantly inhibited VEGF expression of HCC cells and liver tumor tissues. LMWC obviously enhanced and prolonged the deposition of shRNA at the tumor site when LMWC/shRNA complexes were intravenously injected into orthotopic allograft liver tumor-bearing mice. The administration of LMWC/VEGF shRNA complexes by intratumoral or intravenous injection demonstrated more effective suppression of tumor angiogenesis and tumor growth in different HCC models compared with naked shRNA.
  
  
  CONCLUSIONS
  This study demonstrated the feasibility of using LMWC as a potential carrier for RNA interference drugs in liver cancer therapy.
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210093, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Huang
authors.forename: Lei
authors.initials: L
authors.lastname: Dong
authors.forename: Jijun
authors.initials: J
authors.lastname: Chen
authors.forename: Fengbo
authors.initials: F
authors.lastname: Gao
authors.forename: Zhengping
authors.initials: Z
authors.lastname: Zhang
authors.forename: Jiangning
authors.initials: J
authors.lastname: Chen
authors.forename: Junfeng
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy*; D045744:Cell Line, Tumor; D048271:Chitosan / Q000378:metabolism / Q000494:pharmacology*; D017931:DNA Primers / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D018014:Gene Transfer Techniques; D008113:Liver Neoplasms / Q000188:drug therapy*; D051379:Mice; D008813:Mice, Inbred ICR; D008856:Microscopy, Fluorescence; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors*
chemical_list: D017931:DNA Primers; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; D048271:Chitosan