自制纳米级超声微泡造影剂与脂质体基因转染效率的比较

Comparative study on gene transfer efficiency of homemade nanoscale ultrasound microbubbles contrast agent and liposome

目的以GFP为目的基因，比较自制纳米级超声微泡造影剂和脂质体的转染效率，探讨自制纳米级超声微泡造影剂作为基因载体的可行性。方法噻唑蓝（MTT）比色法检测超声微泡浓度对HepG2的细胞毒性；取安全浓度的超声微泡造影剂和脂质体分别与4、8、16μg的PShut—tle—IRES-hrGFP-1质粒结合后转染HepG2细胞，24h后利用荧光显微镜和流式细胞术检测并比较两者的GFP转染效率。结果MTF法显示超声微泡造影剂浓度≤5％时对HepG2细胞生长无明显影响（P〈0．05）；超声微泡造影剂能将GFP基因成功转运到HepG2细胞内并高效表达，微泡造影剂＋8μg质粒组转染效率达（32．61±3．42）％；脂质体＋4μg质粒组的转染效率为（34．12±8．06）％，两组差异无统计学意义（P〉0．05）。结论自制纳米级超声微泡造影剂能成功转运外源DNA进入细胞内，其转染效率与脂质体无明显差异。

Objective Toevaluate the availability of homemade nanoscale uhrasound microbubbles contrast agent as a new gene carrier. Methods MTF was used to measure the cell survival rate to chose the safe concentration of the contrast agent. The mixture of PShuttle-IRES-hrGFP-1 plasmid and microbubbles contrast agent/LipofectamineTM 2000 was transfected into HepG2 cells. Twenty-four h later, the expression of GFP in the cells was observed by fluorescence microscopy,then flow cytomertry was used to evaluate the gene transfer efficiency. Results （ 1 ） ≤5% microbubbles contrast agent had no significant influence on HepG2 growth （P 〈 0.05）. （2） Homemade microbubbles contrast agent could delivery GFP into HepG2 and the genes could be expressed efficiently. The gene transfer efficiency of homemade microbubbles contrast agent was （ 32.61 ± 3.42 ） %, and that of LipofectamineTM 2000 was （ 34.12 ± 8.06） % with the difference being not statistically significant between them （P 〉 0.05 ）. Conclusion Homemade nanoscale microbubbles contrast agent could delivery exogenous DNA into HepG2 cells, and its gene transfer efficiency had no statistically significant difference compared with Lipofectamine^TM 2000.