title: Development of hybrid-type modified chitosan derivative nanoparticles for the intracellular delivery of midkine-siRNA in hepatocellular carcinoma cells.
abstract: BACKGROUND
  Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. Most of the patients with HCC lose the surgical opportunity at the time of diagnosis. Some novel therapeutic modalities, like gene therapy, are promising for the treatment of HCC. However, the success of gene therapy depends on two aspects: efficient gene materials and gene delivery vectors. The present study was to develop new chitosan-based nanoparticles for a midkine-siRNA (anti-HCC gene drug) delivery.
  
  
  METHODS
  The novel gene delivery vector (MixNCH) was synthesized by hybrid-type modification of chitosan with 2-chloroethylamine hydrochloride and N, N-dimethyl-2-chloroethylamine hydrochloride. The chemical structure of MixNCH was characterized by FT-IR and 1HNMR. The cytotoxicity of MixNCH was determined by MTS assay. The gene condensation ability and size, zeta potential and morphology of MixNCH/MK-siRNA nanoparticles were measured. The in vitro transfection and gene knockdown efficiency of midkine by MixNCH/MK-siRNA nanoparticles was detected by qRT-PCR and Western blotting. Gene knockdown effect at the molecule level on the proliferation of HepG2 in vitro was determined by MTS assay.
  
  
  RESULTS
  MixNCH was successfully acquired by aminoalkylation modification of chitosan. The MixNCH could condense MK-siRNA well above the weight ratio of 3. The average size of MixNCH/MK-siRNA nanoparticles was 100-200 nm, and the surface charge was about +5 mV. Morphologically, MixNCH/MK-siRNA nanoparticles were in regular spherical shape with no aggregation. Regarding to the in vitro transfection of nanoparticles, the MixNCH/MK-siRNA nanoparticles reduced MK mRNA level to 14.03%+/-4.03%, which were comparable to Biotrans (8.94%+/-3.77%). MixNCH/MK-siRNA effectively inhibited the proliferation of HepG2 in vitro.
  
  
  CONCLUSIONS
  MixNCH/MK-siRNA nanoparticles could be effective for the treatment of hepatocellular carcinoma.
authors: Huzhou Key Laboratory of Molecular Medicine, Huzhou Central Hospital, Huzhou 313000, China. dlc21@126.com.
authors: Jing
authors: J
authors: Zhong
authors: Hui-Lian
authors: HL
authors: Huang
authors: Jing
authors: J
authors: Li
authors: Fu-Chu
authors: FC
authors: Qian
authors: Li-Qin
authors: LQ
authors: Li
authors: Ping-Ping
authors: PP
authors: Niu
authors: Li-Cheng
authors: LC
authors: Dai
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation; D048271:Chitosan / Q000031:analogs & derivatives / Q000737:chemistry* / Q000633:toxicity; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D018014:Gene Transfer Techniques; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D000078792:Midkine; D053758:Nanoparticles; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013499:Surface Properties; D014162:Transfection
chemical_list: C066254:MDK protein, human; D009414:Nerve Growth Factors; D034741:RNA, Small Interfering; D000078792:Midkine; D048271:Chitosan