title: Multi-target lentivirus specific to hepatocellular carcinoma: in vitro and in vivo studies.
abstract: OBJECTIVE
  We aimed at investigating the effects of the targeted transduction of the Wtp53-pPRIME-miR30-shRNA gene into liver cancer cells, under the mediation of anti-alpha fetoprotein scFv-directed lentivirus, and the inhibitory effect of this system on liver cancer cells.
  
  
  METHODS
  The result of infection was observed by fluorescence microscopy. Polymerase chain reaction and Western blotting were used to demonstrate the successful transduction and transcription of the Wtp53-pPRIME-miR30-shRNA-IGF1R gene. Cell growth was observed via the Cell-Counting Kit-8 Method, and cell apoptosis was detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling. To observe further the effects of AFP-Wtp53-pPRIME-miR30-shRNA-IGF1R therapy in animals, models of BALB-C nude mice bearing subcutaneous human hepatocellular carcinoma were established. The influence of the growth of subcutaneously transplanted tumor, expression of Wtp53 protein, apoptosis, and microvessel formation on the overall level of AFP-Wtp53 pPRIME-miR30-shRNA-IGF1R were also evaluated.
  
  
  RESULTS
  Recombinant lentivirus was successfully constructed, and its functional plaque-forming unit titer was determined as 4.58 × 10(9)plaque-forming units/ml. A positive strand was detected by polymerase chain reaction and Western blotting. Lentiviral construction worked effectively in AFP-positive liver cancer cells. In vitro and in vivo experiments showed that the recombinant lentivirus was more efficacious in inhibiting the proliferation of Hep3B cells.
  
  
  CONCLUSIONS
  The Wtp53-pPRIME-miR30-shRNA gene can be subjected to targeted transduction into liver cancer cells under the mediation of anti-alpha fetoprotein scFv-directed lentivirus. The Wtp53-pPRIME-miR30-shRNA system has targeting ability and lethal effects on liver cancer cells.
authors.affiliation: Department of Hepatobiliary & Pancreatic Surgery, Affiliated Jiangsu Cancer Hospital of Nanjing Medical University, Nanjing 210009, China. zhangyewei@njmu.edu.cn
authors.forename: Ye-Wei
authors.initials: YW
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Niu
authors.forename: Xiang
authors.initials: X
authors.lastname: Lu
authors.forename: Yin-xue
authors.initials: YX
authors.lastname: Yang
authors.forename: He-wei
authors.initials: HW
authors.lastname: Zhao
authors.forename: Xia
authors.initials: X
authors.lastname: He
authors.forename: Guo-Wen
authors.initials: GW
authors.lastname: Yin
authors.forename: Jin-Dao
authors.initials: JD
authors.lastname: Wu
authors.forename: Dong-Liang
authors.initials: DL
authors.lastname: Yan
authors.forename: Jing-feng
authors.initials: JF
authors.lastname: Sun
authors.forename: Jian-fei
authors.initials: JF
authors.lastname: Wen
authors.forename: Ji-feng
authors.initials: JF
authors.lastname: Feng
authors.forename: Huan-zhou
authors.initials: HZ
authors.lastname: Xue
authors.forename: Wan Yee
authors.initials: WY
authors.lastname: Lau
mesh_terms: D000818:Animals; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000098:blood supply / Q000473:pathology / Q000628:therapy*; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008113:Liver Neoplasms / Q000098:blood supply / Q000473:pathology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D035683:MicroRNAs / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D017526:Receptor, IGF Type 1 / Q000032:analysis / Q000235:genetics*; D000509:alpha-Fetoproteins / Q000235:genetics*
chemical_list: C546086:MIRN30b microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; D000509:alpha-Fetoproteins; D017526:Receptor, IGF Type 1