title: Comparative insight into expression of recombinant human VEGF111b, a newly identified anti-angiogenic isoform, in eukaryotic cell lines.
abstract: VEGF-A is a critical growth factor in tumor growth and progression. Two families of VEGF-A isoforms are produced through alternative splicing including VEGFxxx pro-angiogenic and VEGFxxxb anti-angiogenic isoforms. VEGF111b is a new member of the VEGFxxxb family that is induced by mitomycin C and doesn't express in normal conditions. The potent anti-angiogenic properties of VEGF-111b and its remarkable resistance to proteolysis make it an interesting alternative candidate for therapeutic use in all types of cancers. Here, the recombinant VEGF-111b cDNA with insertion of intronic sequence was constructed by using a class IIs restriction enzyme-based method. The recombinant pBud-VEGF111b was transfected into CHO dhfr(-) and HEK 293 cell lines which are currently the standard hosts for the production of candidate therapeutic proteins. Then, the VEGF-111b expression was evaluated in two cell lines using the Real-time PCR. The production of VEGF-111b protein was also investigated here by dot blotting. The VEGF expression was increased about 109 and 185-folds in transfected CHO-dhfr(-) and HEK 293 cells, respectively, in comparison with the un-transfected cells. Dot blotting approach confirmed that both cell lines have successfully produced the VEGF-111b protein.
authors.affiliation: Division of Genetics, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Electronic address: f.dehghanian68@gmail.com.
authors.forename: Fariba
authors.initials: F
authors.lastname: Dehghanian
authors.affiliation: Division of Genetics, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Electronic address: z.hojati@sci.ui.ac.ir.
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Hojati
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D045744:Cell Line, Tumor; D017931:DNA Primers; D006801:Humans; D020033:Protein Isoforms / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D020033:Protein Isoforms; D042461:Vascular Endothelial Growth Factor A
keywords: Cancer; Dot blotting; Real-time PCR; VEGF-111b