阳性脂质体介导反义寡聚核苷酸转染HeLa细胞的效果研究

THE TRANSFECTION EFFICIENCY OF OLIGONUCLEOTIDES TRANSFECTION MEDIATED BY CATIONIC LIPOSOMES IN HELA CELLS

采用FITC标记的未经修饰的和经过修饰的两种19-mer反义寡聚核苷酸序列(ODN19和S-ODN19)作为转染物质,用流式细胞技术(FCM)研究比较几种常用阳性脂质体介导的寡聚核苷酸转染HeLa细胞的效果及适宜的转染时间。未经化学修饰的ODN19转染结果显示,LipofectAmine和DM-RIE-C增强转染的作用相对较强,而其他两种脂质体的作用并不明显。对于经过修饰的S-ODN19转染而言,四种阳性脂质体均具有增强S-ODN19转染作用,但以LipofectAmine的效果最为明显,其转染效果(FITC均值为5203.11)为无脂质体介导对照的数十倍。四种阳性脂质体的增强转染作用排序为:LipofectAmine>FuGENE6>Lipofectin>DM-RIR-C。另外,在用FuGENE6介导寡聚核苷酸转染时,采用4小时转染时间可获较好转染效果。

The enhance effect of oligonucleotide transfection mediated by cationic liposomes (Lipofectin, LipofectAmine, FuGENE6 and DM-R1E-C) were investagated using flow cytometry in Hela cells. Two 19-mer molecules, phosphodiester oligonucleotide (ODN19) and phosphorohieoate oligonucleotides (SODN19), were employed, which are labeled with Fluorescein isothiocyanate(FITC) and have the same sequence. The results show that, for the unmodified ODN19 molecules, LipofectAmine and DMRIE-C can obviously improve the transfection, but no improving effect were observed in other two liposome. For the S-ODN19 transfection, all of the 4 calionic liposomes possess the observed ability to enhance the transfection, of which the LipofectAmine was more efficient than three others. The mean FITC density (5203.11) of LipofectAmine mediated transfection was several decades times comparable to the control cultures without mediated liposome. The improving effect of liposomes mediated transfection was ranked as LipofectAmine > FuGENE6 > DMRIE-C > lipofectin. Meanwhile. The experiment also suggested that, when transfection procedure was performed for about 4 hours, a high efficiency can be expected in FuGENE6 mediated oligonucleotides transfection.