Oh Binna, Han Jaesik, Choi Eunji, Tan Xiaonan, Lee Minhyung
BK21 Plus Future Biopharmaceutical Human Resources Training and Research Team, Department of Bioengineering, College of Engineering, Hanyang University, Seoul, 133-791, Republic of Korea.
J Pharm Sci. 2015 Apr;104(4):1461-9. doi: 10.1002/jps.24363. Epub 2015 Jan 28.
Bevacizumab (Avastin) is an angiogenesis inhibitor used as a treatment for various cancers. In this study, the combination therapy of Avastin and glioblastoma-specific thymidine kinase gene [pEpo-NI2-SV-herpes simplex virus thymidine kinase(HSVtk)] was evaluated in a glioblastoma animal model. The R7L10 peptide was used as a gene carrier of pEpo-NI2-SV-HSVtk. Gel retardation assays confirmed that R7L10 formed stable complexes with pEpo-NI2-SV-HSVtk. R7L10 protected DNA from nuclease digestion. R7L10 had lower transfection efficiency than polyethylenimine (PEI; 25 kDa). However, the in vitro and in vivo toxicity assays showed that R7L10 had lower cytotoxicity than PEI, suggesting that R7L10 is safer than PEI. For the combination therapy, Avastin was injected intravenously and the pEpo-NI2-SV-HSVtk/R7L10 complexes were injected intratumorally in the glioblastoma animal model. Tumor growth was most effectively inhibited by the combination therapy of Avastin and the gene. The immunostaining results confirmed that the HSVtk genes were expressed in the groups with the pEpo-NI2-SV-HSVtk/R7L10 complex. The terminal deoxynucleotidyl transferase dUTP nick end labeling assay showed a higher level of apoptotic cells in the combination group than the pEpo-NI2-SV-HSVtk/R7L10 complex or Avastin group. In conclusion, the combination of Avastin and the glioblastoma-specific HSVtk gene has a higher antitumor effect than single therapy of Avastin or HSVtk after intratumoral administration in glioblastoma animal model.
贝伐单抗（阿瓦斯汀）是一种血管生成抑制剂，用于治疗多种癌症。在本研究中，在胶质母细胞瘤动物模型中评估了阿瓦斯汀与胶质母细胞瘤特异性胸苷激酶基因[ pEpo - NI2 - SV -单纯疱疹病毒胸苷激酶（HSVtk）]的联合治疗。R7L10肽用作pEpo - NI2 - SV - HSVtk的基因载体。凝胶阻滞试验证实R7L10与pEpo - NI2 - SV - HSVtk形成稳定复合物。R7L10可保护DNA免受核酸酶消化。R7L10的转染效率低于聚乙烯亚胺（PEI；25 kDa）。然而，体外和体内毒性试验表明，R7L10的细胞毒性低于PEI，这表明R7L10比PEI更安全。对于联合治疗，在胶质母细胞瘤动物模型中静脉注射阿瓦斯汀，并瘤内注射pEpo - NI2 - SV - HSVtk / R7L10复合物。阿瓦斯汀与该基因的联合治疗最有效地抑制了肿瘤生长。免疫染色结果证实，在含有pEpo - NI2 - SV - HSVtk / R7L10复合物的组中表达了HSVtk基因。末端脱氧核苷酸转移酶dUTP缺口末端标记试验显示，联合组中的凋亡细胞水平高于pEpo - NI2 - SV - HSVtk / R7L10复合物组或阿瓦斯汀组。总之，在胶质母细胞瘤动物模型中瘤内给药后，阿瓦斯汀与胶质母细胞瘤特异性HSVtk基因的联合具有比阿瓦斯汀或HSVtk单一治疗更高的抗肿瘤作用。
