title: [Construction, expression and targeting therapeutic of single-chain immunotoxin against hepatocellular carcinoma].
abstract: OBJECTIVE
  To obtain high therapeutic effect and low toxicity single-chain immunotoxin against hepatocellular carcinoma (HCC).
  
  
  METHODS
  Human mutant tumor necrosis factor-alpha (mTNFalpha) was linked with the 3' end of humanized single-chain Fv against HCC (hscFv25) in pGEX4T-1 vector. The anti-HCC immunotoxin was expressed in Escherichia coli and identified by western blot. The primary tumor regression trial in nude mice bearing HCC was evaluated the targeting therapeutic value of hscFv25-mTNFalpha. The tumor tissues were stained by immunohistochemical with TNFalpha antibody.
  
  
  RESULTS
  The expression of single-chain immunotoxin hscFv25-mTNFalpha was 12% of total bacteria proteins. The tumor regression trials of hscFv25-mTNFalpha showed 5/5 effective. It had 2/5 completely remission and 3/5 partly remission. The therapeutic result of hscFv25-mTNFalpha was better than that of mTNFalpha (F=8.70, 0.05). The HCC tissue treated by hscFv25-mTNFalpha expressed TNFalpha positive reaction. The positive granule mainly existed in HCC cytoplasm.
  
  
  CONCLUSIONS
  The single-chain immunotoxin hscFv25-mTNFalpha has high therapeutic effect and low toxicity. It has potentialities for clinical application.
authors.affiliation: Department of Pathology, Xijing Hospital, Fourth Military Medical University, Xi'an 710033, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Yan-fang
authors.initials: YF
authors.lastname: Liu
authors.forename: Shou-jing
authors.initials: SJ
authors.lastname: Yang
authors.forename: Qing
authors.initials: Q
authors.lastname: Qiao
authors.forename: Hong
authors.initials: H
authors.lastname: Cheng
authors.forename: Fu-cheng
authors.initials: FC
authors.lastname: Ma
authors.forename: Wan-lu
authors.initials: WL
authors.lastname: Chen
mesh_terms: D000818:Animals; D007128:Immunoglobulin Fragments / Q000627:therapeutic use*; D007150:Immunohistochemistry; D000922:Immunotoxins / Q000627:therapeutic use*; D008114:Liver Neoplasms, Experimental / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D014409:Tumor Necrosis Factor-alpha / Q000627:therapeutic use*
chemical_list: D007128:Immunoglobulin Fragments; D000922:Immunotoxins; D014409:Tumor Necrosis Factor-alpha; C055966:immunoglobulin Fv