脂质体介导的端粒酶反义寡核苷酸在人膀胱癌EJ细胞中的转染率及稳定性

目的探讨端粒酶反义寡核苷酸（asODN）在脂质体介导下在入膀胱癌EJ细胞中的转染率、分布特点及稳定性。方法合成针对端粒酶RNA模板区的asODN，并行全硫代修饰或不修饰，5-FITC标记，脂质体介导转染人膀胱癌EJ细胞后分别于15、30min、1h以及以后每小时，在电镜下观察asODN在细胞中的表达，分布特点及稳定性。结果在脂质体介导下，未修饰型asODN转染细胞后15min开始表达，1～3h稳定表达于细胞中，表达率15％～20％，4h减退；修饰型asODN 15min开始表达，8h有56％～80％的高表达，12h开始消散。结论脂质体是一种有前途的膀胱癌非病毒基因治疗载体。

In vitro transfection efficiency of telomerase antisense oligodeoxynucleotide mediated by liposome in human bladder cancer cell line EJ and its stability

Objective To investigate the transfecfion efficiency of telomerase antisense oligodeoxynucleotide related by liposome in human bladder cancer cell line EJ and its stability.Methods Phosphorothiate and unmodified telomerase antisense oligodeoxynucleotides with fluorescein 5'-isothio- cyanate(5'-FITC)were mediated by liposome to human bladder cancer cell line EJ.Intracellular distri- bution of asODN was observed under fluorescence microscopy in fixed cells.Results In the absence of li- posome,the asODN localized to discrete structure in cytoplasm of the cell,resulting in a punctuate fluores- cence pattern in 4 h after treatment.In the presence of liposome,celler fluorescence was markedly in- creased.The transfection efficiency was 56%-80%,notably higher than that in the absence of liposome, 15%-20%.Conclusion Cationic lipids increase the amount and alter intracellular distribution of asODN in bladder cancer cell line EJ,which is more stably expressed in cells.Liposome is potential as non-viral carry for gene therapy.