目的：优化鱼精蛋白 脂质体 寡聚核苷酸(oligodeoxynuleotides, ODN)复合体（liposome-protamine-ODN,LPD）法以提高SKOV3细胞的转染效率，并探讨该转染法对细胞体外增殖能力的影响。方法：参照文献设计核转录因子 κB（NF-κB）诱捕物ODN和对照物Scrambled ODN，并用FITC标记。LPD表面+/-电荷比例设计为1，2，4，6，并按转染液中是否含有小牛血清分成2组。用荧光光度计、流式细胞仪测定转染效率。用MTT法观察LPD法对SKOV3细胞在体外增殖的影响。结果：SKOV3细胞经LPD法转染，＋/－电荷比例为4时转染效率最高，达96.41％，而无鱼精蛋白-脂质体的裸露ODN细胞转染效率为12.87％,两者差异有统计学上的高度显著性意义。培养液中10％小牛血清对转染效率无明显影响。结论：适宜电荷比例的LPD法是一种不受血清影响的高效率转染方法，有望用于体内转染。

Objective: To observe the transfer efficiency of protamine-DOTAP/Chol liposome-oligodeoxynuleotides complex to SKOV3 cells. Methods: Synthesized NF-κB decoy ODNs or scrambled ODNs labeled by FITC on the 5′ends was mixed with rationic DOTAP/cholesteral liposome and protamine, making the charge ratio of positive:negative 4. SKOV3 cells were transfected by the complex(1 μmol/L) and the efficiency were measured by flow cytometry(FCM) and fluorescence meter. The direct effect of NF-κB decoy ODNs transfer on the growth of SKOV3 cell line in vitro was measured by MTT method. Results: The efficiency of LPD transferto SKOV3 cell line was 96.41%, significantly higher than that of naked DNA transfer which was 12.86%.The growth of SKOV3 cell line was not affected by 48h incubation after NF-κB decoy ODNs or scrambled ODNs transfection.Conclusion: LPD method is the one with high transfer efficiency not affected by NBS(newborn bovine serum)with possibility of use in viv.

R730.3

上海市卫生局“百人计划”基金98BR040赞助