【摘要】 目的 探讨Survivin反义核酸技术诱导乳腺癌细胞凋亡的作用及效果。方法 本实验共分6组，分为以脂质体介导反义寡核苷酸组(ASODN/Lip)、无义寡核苷酸组(NODN/Lip)及RPMI 1640培养液的空白对照组(Lip)，转染人乳腺癌MCF-7细胞系，应用Hoechst 33258/PI双重染色观察MCF-7细胞的形态学改变，透射电镜观察细胞超微结构，流式细胞仪检测细胞周期和细胞凋亡的变化，DNA凝胶电泳观察凋亡情况，研究Survivin反义寡核苷酸对MCF-7乳腺癌细胞的生长抑制作用。结果 Hoechst 33258/PI染色及透射电镜观察可见转染后的细胞结构呈凋亡样改变；流式细胞仪检测见转染后细胞凋亡显著增加，并出现G2/M期阻滞现象；DNA凝胶电泳在600ng/ml，800ng/ml ASODN/Lip组的电泳图谱上呈现出明显的“梯状”现象。结论 凋亡抑制基因Survivin表达下调对乳腺癌细胞的生长抑制作用主要是通过诱导细胞发生凋亡以及阻止有丝分裂发生在G2/M阻滞期来实现，Survivin靶向反义核酸技术可能成为乳腺癌基因治疗的一种有效方法。

    【关键词】 乳腺癌；Survivin基因；反义核酸技术；凋亡

    Cells apoptosis induced by survivin antisense oligonucleotide

    DUAN Ti-de,LIU De-quan, YANG Ce-yao.

    Department of General Surgery,The First Affiliated Hospital of Kunming Medical College,Kunming 650032,China

    【Abstract】 Objective To explore the effect of cells apoptosis on human breast cancer after survivin was blocked with the antisense technique.Methods The experiments were divided into six groups. Antisense Oligonucleotide (ASODN), nonsense oligonucleotide (NODN) was transfected in human breast cancer MCF-7 cell line and RPMI 1640 culture medium (normal control). The study used Hoechst 33258/PI,electron microscope, flow cytometer and in gel electrophoresis of DNA to observe the cells apoptosis.Results The cells apoptosis after transfection were observed by Hoechst 33258/PI,electron microscope and in gel electrophoresis of DNA.The result of flow cytometer showed the cells apoptosis after transfection and the blocking phenomena appeared in G2/M period.Conclusion The restriction by the reduction of survivin expression in breast cancer cells is mainly functioned through its induction of cells apoptosis and prevention of mitosis at G2/M period. Survivin targeting therapy can be used as an important method in breast cancer treatment. ......