PEI在动物皮肤组织基因转化中的应用研究

Study on the Application of PEI for Gene Transfer in Mouse Skin Tissue

摘要:

用低分子量的聚乙亚胺（PEI）开发一种新的非病毒基因转染系统，为基因在皮肤组织中的有效转染提供一种可靠、廉价的方法。将带有绿色荧光蛋白报告基因 (gfp) 的真核表达质粒与阳离子聚合物聚乙亚胺结合，用肝癌细胞株CM7721试验，研究其转染效率及可能引起的细胞毒性；进一步转染小鼠皮肤组织，研究转染基因的表达位置及持续表达时间。结果发现，低分子量PEI介导的细胞转染效率最高可达55%，转染效率与PEI结构无关（P>0.05），但是随着PEI分子量的增加，其转染活性略有下降。同时，随着分子量的增加，PEI对细胞的毒性也相应加大；小鼠皮肤转染实验显示，转染24h后，gfp即可在皮肤组织的毛囊、汗腺、皮脂腺等处高效表达，表达可持续7～9d；进一步对皮肤用氮酮、维甲酸处理后，gfp可在皮肤组织的颗粒层细胞中高效表达。PEI是一种高效、有用的非病毒基因转染载体，能够在体外培养的动物细胞及动物皮肤组织中进行基因转移，这对皮肤疾病的基因治疗具有潜在的应用价值。

Abstract:

A reliable, low-cost, and highly efficient nonviral gene delivery system using lower molecular weight polyethylenimine (LMW-PEI) is provided. LMW-PEI was linked to an expressing plasmid with green fluorescence protein gene (gfp), the transfection activity mediated by PEIs were examined in the CM7721 cell line and the skin tissue of mouse, respectively. The cytotoxicity of PEIs, the localization and continuance time of gfp expressed in the skin tissue of mouse were also studied. Results showed that the transfection rate of gfp mediated by LMW-PEI in the CM7721 cell line was about 55%. However, with the increasing PEI molecular weight, the cytotoxicity of PEI increased, but its transfection activity decreased. The tissue transfection results showed that LMW-PEI induced a significant expression of the gfp in the cells of hair vesicle and sweat gland of mouse skin tissues following transfection of 24 h, and the expression of gfp lasted 7～9 d. When the tissue of mouse was treated with retinoic acid and nitrogenous ketone, respectively, gfp was transferred to the granule layer of mouse skin tissue. The LMW-PEI described here is a new, highly efficient vector; it would be a useful nonviral vector for gene delivery technology.