脂质体介导的c-erbB-2反义寡核苷酸转染小鼠乳腺癌TM40D细胞的效率及作用

目的 探讨阳离子脂质体介导的c-erbB-2反义寡核苷酸(ODNs)转染小鼠乳腺癌TM40D细胞的效率及其对细胞增殖、凋亡的影响。方法 应用流式细胞仪及荧光显微镜观察反义ODNs在1、2、4、6h的转染效率以及在细胞内的分布。应用四甲基偶氮唑蓝比色分析法(MTT)检测反义ODNs对细胞增殖的影响;流式细胞仪检测对细胞凋亡的影响。结果 脂质体可促进反义ODNs进入细胞内,无脂质体介导的反义ODNs转染细胞的效率随转染时间延长逐步增加,而脂质体介导的反义ODNs转染细胞4h效率达到高峰72．23％,6h维持在高水平。脂质体可使反义ODNs在细胞内有效分布,进入细胞核。MTT法检测结果表明c-erbB-2反义ODNs抑制细胞增殖,抑制率为50．24％。流式细胞仪检测结果表明c-erbB-2反义ODNs促进细胞凋亡,正常细胞组凋亡率为9．13％,脂质体组为9．29％,脂质体 反义ODNs组为38．50％。结论 脂质体可促进反义ODNs进入细胞及在细胞有效分布,c-erbB-2反义ODNs可抑制乳腺癌TM40D细胞的增殖及促进癌细胞凋亡。

Optimal effects of liposome on antisense oligodeoxynucleotides uptake by mouse breast cancer TM40D cells and the effects of antisense oligodeoxynucleotides targeting c-erbB-2 on cell proliferation and apoptosis

Objective To study the optimal effects of liposome on antisense oligodeoxynucleotides targeting c erbB 2 uptake by mouse breast cancer TM40D cells and effects of AS ODNs on cell proliferation and apoptosis. Methods Flow cytometric analysis and fluorescence microscope were used to study the transfection ratio of AS ODNs in mouse breast cancer TM40D cells with/without liposome in 1?2?4?6 hour and the distribution of AS ODNs in cells. the effects of ODNs on cell proliferation and activation of apoptosis were examined by MTT assay and flow cytometry. Results Liposome could promote AS ODNs uptake by mouse breast cancer TM40D cells, flow cytometric analysis revealed that the time of the optimal effects was 4 hours and the percentage of FAM positive cells reached 72 23%; the ratio maintained at a relatively high level at 6h. Also liposome facilitated the entrance of AS ODNs into nucleus. AS ODNs restrained the proliferation of mouse breast cancer TM40D cells ,and restraint rate was 50 24%. Meanwhile AS ODNs promoted cell apoptosis. Flow cytometry analysis revealed that antisense ODNs increased cell apoptosis by38.50%, compared with cultured cells group 9.13% and liposome group 9 29%. Conclusions Liposome could facilitate AS ODNs to enter cells and their nucleus. AS ODNs of C erbB 2 restrained the proliferation of mouse breast cancer TM40D cells and promoted apoptosis.