FGF receptor-mediated gene delivery using ligands coupled to polyethylenimine.
To obtain new nonviral vectors with high gene delivery efficiency and special cell targeting ability, an attractive strategy is to link ligands to polyethylenimine (PEI). Fibroblast growth factor receptors (FGFRs) are highly expressed on a variety of human cancer cells and are potential targets for cancer gene therapy. In this study, the peptides NH2-Met-Gln-Leu-Pro-Leu-Ala-ThrGly-Gly-Gly-Cys-COOH (MC11) which have been proved to combine specially with the FGFR on cell membrane are coupled to PEI using N-Succinimidyl-3-(2-pyridyldithio) propionate (SPDP) as a linker with different molar ratios (1 : 0.3, 1 : 0.75, 1 : 1.5, and 1 : 3.0) and the new polymer PEI-MC11 is verified by a series of physicochemical methods including 1H-NMR and FTIR. The agarose gel electrophoresis assay, particle size test, zeta potential test, and electron microscope observation show that PEI-MC11 can efficiently condense plasmid DNA into nanoparticles with about 200 nm in diameter and with positive surface charge at the suitable N/P ratio. The MTT assay suggests the decreased toxicity of the polymers. The results of the gene delivery efficiency in vitro show that PEI-MC11/pDNA polyplexes have significantly greater transgene activity than PEI/pDNA in COS-7 and HepG2 cells which express FGFR positively, while no such effect is observed in PC3 cells which have negative FGFR. The enhanced gene delivery efficiency of PEI-MC11 can be blocked by the co-culture of free peptides MC11 before the gene delivery procedure. The synthesized nonviral vector based on PEI with the targeting peptides MC11 for binding FGFR has improved efficiency of gene delivery and targeting specificity in FGFR positive cells. It may have potential application in cancer gene therapy.
Li
Da
D
Institute of Immunology, Zhejiang University Hangzhou, 310058, PR China.
Yu
Hai
H
Huang
Hongliang
H
Shen
Fenping
F
Wu
Xiaoyuan
X
Li
Jingzhong
J
Wang
Jianli
J
Cao
Xuetao
X
Wang
Qingqing
Q
Tang
Guping
G
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004278:DNA, Superhelical / Q000737:chemistry*; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000633:toxicity; D010957:Plasmids / Q000737:chemistry; D011094:Polyethyleneimine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000633:toxicity; D017468:Receptors, Fibroblast Growth Factor / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared
D004278:DNA, Superhelical; D008024:Ligands; D010455:Peptides; D017468:Receptors, Fibroblast Growth Factor; C524332:polyethylenimine-N-succinimidyl-3-(2-pyridyldithio)propionyl-MC11; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine