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

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,24 h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 MTT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+4 μg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.

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

Objective To evaluate the availability of homemade nanoscale ultrasound microbub-bles contrast agent as a new gene carrier. Methods MTT 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 mi-crobubbles contrast agent/Lipofectamine~(TM) 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 micro-bubbles contrast agent was (32.61 ± 3.42) %, and that of Lipofectamine~(TM) 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 trans-fer efficiency had no statistically significant difference compared with Lipofectamine~(TM)2000.