Development of gene delivery system using nanocoloid and its application for gene targeting therapy

对恶性肿瘤和先天性基因疾病进行了基因治疗试验；然而，基因治疗在人体应用中存在着效率低、风险大等问题，目前还没有形成理想的基因治疗方法。我们设计了将表达质粒包裹在小颗粒内，配体肽包裹在小颗粒外的纳米胶体，用于开发特定的基因传递系统。我们用Western blot法检索了口腔鳞癌细胞系（9.22,HSC2, HSC3, HSC4, Ca SAS）中EGFR的表达，所有细胞系均表达EGFR。我们用BireCore x检测了EGFR与合成EGF肽的结合亲和力，结果表明合成肽与EGFR结合。接下来，我们研究了纳米胶体对口腔癌细胞系的诱导作用。转染效率约为30%，我们的目标是提高效率。我们接下来检查了肾上腺髓质素拮抗剂的作用，它具有抑制血管生成的能力。具有抑制肾上腺髓质素序列的载体能抑制裸鼠移植瘤的生长。这些结果提示针对肿瘤环境的特异性基因传递可能成为实体癌的治疗药物。

Trials of gene therapies have been carried out for malignant tumors and congenital gene disorders,; however, there are problems such as the low efficiency and risk for applying human being, and ideal methods of gene therapy are not yet developed.. We designed nanocoloid that enclosed expression plasmid inside and, ligand peptide outside of small particle for development of a specific gene delivery system. When We searched expression of EGFR in oral squamous cell carcinoma cell lines (9.22, HSC2, HSC3, HSC4, Ca SAS) by Western blot, and all cell lines expressed EGFR We examined the binding affinity of EGFR and synthesized EGF peptide by BireCore X. As a result, it became clear that synthesized peptide bound to EGFR. Next, we examined the induction effect of nanocoloid to oral carcinoma cell lines. The efficiency of transfection was approximately 30%, and we aimed to improve the efficiency. We next examined the effects of adrenomedullin antagonist that has the ability to inhibit angiogenesis. The vector that has the sequence for inhibitory effects on aderenomedullin reduced the Growth of tumor xenografts in nude mice.These results imply that specific gene delivery for tumor environment could be the therapeutic agents for solid cancers.