title: Expression, purification and characterization of recombinant toxins consisting of truncated gastrin 17 and pseudomonas exotoxin.
abstract: Gastric cancer is a major cause of mortality and morbidity around world. However the effectiveness of the current approaches to the diagnosis and treatment of gastric cancer is limited. Recombinant targeted toxins may represent a novel direction of cancer therapy. In this study, we aimed to explore whether recombinant toxins fused with the truncated forms of G17 could target to kill cancer cells by recognizing CCK2R. Four recombinant Pseudomonas toxins PE38 fused with the forward or reverse truncated forms of G17 (G14 and G13) were successfully constructed, expressed, and purified. Their characteristics were further analyzed by SDS-PAGE, western blot and indirect immunofluorescence assay. The cytotoxicity assay demonstrated that only reversely fused recombinant toxins rG14PE38 and rG13PE38 exhibited certain toxicity on several cancer cell lines, and a competition assay indicated that the binding of the reverse gastrin-endotoxin to CCK2R (+) cells may be mediated by interaction between gastrin/gastrin-like and CCK2R.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Feng
authors.forename: Xi-Lin
authors.initials: XL
authors.lastname: Liu
authors.forename: Shi-Ying
authors.initials: SY
authors.lastname: Lu
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Ren
authors.forename: Yan-Song
authors.initials: YS
authors.lastname: Li
authors.forename: Pan
authors.initials: P
authors.lastname: Hu
authors.forename: Quan
authors.initials: Q
authors.lastname: Wang
authors.forename: Weihua
authors.initials: W
authors.lastname: Tong
authors.forename: Dong-Ming
authors.initials: DM
authors.lastname: Yan
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.forename: Song
authors.initials: S
authors.lastname: Zhang
authors.forename: Wen
authors.initials: W
authors.lastname: Jin
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education / Institute of Zoonosis / College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China. zsliu1959@163.com.
authors.forename: Zeng-Shan
authors.initials: ZS
authors.lastname: Liu
mesh_terms: D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005098:Exotoxins / Q000235:genetics / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D005755:Gastrins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009369:Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000737:chemistry; D043887:Receptor, Cholecystokinin B / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D014118:Toxins, Biological / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D005098:Exotoxins; D005755:Gastrins; D043887:Receptor, Cholecystokinin B; D011994:Recombinant Proteins; D014118:Toxins, Biological; C013550:gastrin 17