摘要
目的研究声振脂质微泡(PGM)作为反义寡核苷酸传递系统的可行性。方法应用PGM将结合荧光蛋白的反义核苷酸片段ZL转染到乳腺癌细胞SK-BR-3中,考察不同因素如:ZL浓度、PGM浓度、超声持续时间和机械指数(M I)等对ZL转染率及细胞存活率的影响。考察PGM的结果,并与其他脂质载体如lipofectam ine和脂质体的转染效果比较。应用荧光显微镜观察转染率,碘化丙锭(PI)染色观察细胞存活率。结果考察因素中,PGM浓度,超声持续时间和M I对ZL转染率和细胞存活率影响较大。最佳超声条件为:PGM浓度为2%,超声持续时间为30 s和M I为1.0,在此条件下转染率为78%±10%,比未超声的PGM(4.0%)和lipofectam ine(4.3%)的转染率增加了约18倍,而细胞存活率相近。结论在适宜条件下,声振脂质微泡可以有效促进体外反义核苷酸的转染。
Aim To investigate the feasibility of transfer antisense oligodeoxynucleotides （AS-ODNs） by the phospholipids-based gas-filled microbubbles （PGM） under ultrasound activation. Methods An antisense oligodeoxynucleotides sequence ZL combined with luciferase reporter plasmid was used. A breast cancer cell line SK-BR-3 was exposed to different conditions to investigate the effects of such factors as ZL concentration, PGM concentration, mechanical index （MI） and ultrasound exposure duration on transfection efficiency and cell viability. The transfection efficiency and cell viability by other lipid vectors such as lipofectamine and liposome were also tested, whose results were comparied with that of PGM. Transfection efficiency was detected by fluorescence microscopy. Cell viability was verified by PI （propidium iodide） assay. Results Among the factors tested, ultrasound exposure duration, MI and PGM concentration had obvious impacts on transfection efficiency and cell viability. The results showed that the optimal ultrasound condition was the exposure to ultrasound at MI 1.0 for 30 s with 2% PGM concentration, which gave an overall transfection efficiency of 780/6 ±10% , increased nearly 18 folds over the transfection by PGM （4.0%） or lipofectamine （4. 3% ） without ultrasound. Under same ultrasound conditions, different vectors showed significant difference in transfection efficiency while there are similar results in cell viability. Conclusion Under proper ultrasound conditions, PGM can markedly enhance AS-ODNs transfection efficiency.
出处
《药学学报》
CAS
CSCD
北大核心
2006年第9期899-904,共6页
Acta Pharmaceutica Sinica
关键词
超声
微泡
反义核苷酸
基因转染
ultrasound
microbubbles
antisense oligodeoxynucleotides
gene transfection
