title: Design, synthesis and evaluation of VEGF-siRNA/CRS as a novel vector for gene delivery.
abstract: Small interfering RNA (siRNA) delivery is a prospective method in gene therapy, but it has application limitations such as negative charge, water solubility and high molecular weight. In this study, a safe and efficient nano-vector, CRS, was designed and synthesized to facilitate siRNA delivery. Physical and chemical properties of VEGF-siRNA/CRS were characterized by methods including scanning electron microscopy (SEM), transmission electron microscopy, zeta potential (ζ) measurement, drug-releasing rate measurement, gel electrophoresis and confocal microscopy. The biological activities were evaluated using cell viability assay, gene-silencing efficacy assay in vitro, real-time polymerase chain reaction, enzyme-linked immunosorbent assay (ELISA) and antitumor tests in vivo. The mean nanoparticle size of VEGF-siRNA/CRS was 121.4±0.3 nm with positiveζpotential of 7.69±4.47 mV. The release rate of VEGF-siRNA from VEGF-siRNA/CRS was 82.50% sustained for 48 h in Tris-ethylenediaminetetraacetic acid buffer (pH 8.0). Real-time polymerase chain reaction was used to analyze the efficiency of the transfection, and the result showed that VEGF mRNA expression had been knocked down by 82.36%. The expression of VEGF protein was also recorded to be downregulated to 14.83% using ELISA. The results of cytotoxicity measured by Cell Counting Kit-8 assay showed that VEGF-siRNA/CRS had significant inhibitory effect on HeLa cells. The results of antitumor assays indicated that VEGF-siRNA/CRS exhibited tumor cell growth inhibition in vivo. The results demonstrated that VEGF-siRNA could be delivered and transported by the designed carrier, while siRNA could be released constantly and led to an increasing gene-silencing effect against VEGF gene. In conclusion, VEGF-siRNA/CRS is a promising carrier for siRNA delivery, and further studies are warranted.
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Zhao
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China.
authors.forename: Xueyun
authors.initials: X
authors.lastname: Jiang
authors.affiliation: School of Chemical Biology and Pharmaceutical Sciences, Capital Medical University, Beijing, China.
authors.forename: Chunying
authors.initials: C
authors.lastname: Cui
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use; D002470:Cell Survival; D015536:Down-Regulation; D016503:Drug Delivery Systems; D065546:Drug Liberation; D015316:Genetic Therapy; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D053758:Nanoparticles / Q000737:chemistry* / Q000633:toxicity; D009369:Neoplasms / Q000235:genetics / Q000628:therapy*; D009842:Oligopeptides / Q000737:chemistry / Q000633:toxicity; D010316:Particle Size; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; D009842:Oligopeptides; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; C046617:arginyl-glycyl-aspartyl-serine
keywords: CRS; VEGF; antitumor; biophysical characteristics; gene silencing; protein expression; siRNA delivery; tumor therapy