In vivo growth inhibitory effect of iterative wild-type p53 gene transfer in human head and neck carcinoma xenografts using glucosylated polyethylenimine nonviral vector
In vivo growth inhibitory effect of iterative wild-type p53 gene transfer in human head and neck carcinoma xenografts using glucosylated polyethylenimine nonviral vector
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DOI：
10.1038/sj.cgt.7700485
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发表时间：
2002-08-01
影响因子：
6.4
通讯作者：
Guillemin, F
中科院分区：
文献类型：
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作者：
Dolivet, G;Merlin, JL;Guillemin, F
聚乙烯亚胺（PEI）衍生物是用于基因转移的聚阳离子非病毒载体。先前在头颈癌细胞中体外获得的结果表明，与未取代的PEI相比，葡糖基化PEI产生更高的基因转移效率和更长的转基因表达。使用葡萄糖基化PEI，成功地实现了p53基因转移，随后恢复P53蛋白表达并诱导自发性凋亡。本研究报告了在人头颈部鳞状细胞癌异种移植小鼠中获得的体内数据，使用生物素化PEI和组织化学分析，发现载体在肿瘤组织的增殖细胞中扩散，保留坏死区域。在由非增殖、成熟分化细胞组成的角化区域内未观察到扩散。利用绿色荧光蛋白（GFP）转染和荧光显微镜观察，转基因表达主要在含有增殖细胞的肿瘤周围。GFP表达在肿瘤深度内较低。然后使用荧光素酶作为报告基因测定定量转基因表达动力学。在肿瘤内注射葡糖基化PEI/DNA复合物后48小时达到最大转基因表达。两次瘤内注射后48小时达到最高的基因转移效率。转染野生型p53后，在接受每周两次重复的葡糖基化PEI/DNA复合物的肿瘤内注射的荷瘤小鼠中观察到肿瘤生长抑制。使用相同的时间表在连续治疗下维持肿瘤生长抑制。在所有实验中，未观察到明显的毒性。本研究结果证明了利用葡萄糖基化聚乙烯亚胺进行非病毒基因转移的可行性和肿瘤生长抑制效力。
Polyethylenimine (PEI) derivatives are polycationic nonviral vectors for gene transfer. Previous results achieved in vitro in head and neck cancer cells demonstrated that glucosylated PEI yields higher gene transfer efficiency and longer transgene expression than unsubstituted PEI. Using glucosylated PEI, p53 gene transfer was successfully achieved with subsequent recovery of P53 protein expression and induction of spontaneous apoptosis. The present study reports in vivo data achieved in human head and neck squamous cell carcinoma xenografted mice, Using biotinylated PEI and histochemistry analysis, the vector was found to diffuse in the proliferating cells of the tumor tissue, sparing necrotic areas. No diffusion was observed inside keratinized area composed of nonproliferating, mature differentiated cells. Using green fluorescent protein (GFP) transfection and fluorescence microscopy, the transgene expression was mainly observed at the periphery of the tumor containing proliferating cells. GFP expression appeared lower inside the tumor depth. Quantitative transgene expression kinetics was then determined using luciferase as reporter gene. The maximal transgene expression was achieved 48 hours after intratumoral injection of glucosylated PEI/DNA complexes. The highest gene transfer efficacy was achieved 48 hours after two intratumoral injection. After transfection of wild-type p53 tumor growth inhibition was observed in tumor-bearing mice receiving intratumoral injection of glucosylated PEI/DNA complexes repeated twice weekly. Tumor growth inhibition was maintained under continuous treatment using the same schedule. In all experiments, no noticeable toxicity was observed. The present results demonstrate the feasibility and the tumor growth inhibition potency of nonviral gene transfer using glucosylated polyethylenimine.