靶向Survivin的反义寡核苷酸对肿瘤细胞增殖的抑制作用

 Survivin是新近克隆的一种凋亡抑制蛋白 (IAP)家族成员 ,在几乎所有肿瘤组织中特异性表达 ,而在正常成年终末分化组织中低表达甚至不表达 .采用四唑盐 (MTT)比色实验法比较 2 0条抗人survivin反义寡核苷酸对HeLa细胞增殖的抑制效果 ,并从中筛选效果显著的反义寡核苷酸 ,在体外水平进一步验证其抑制survivin表达的能力 .在用 4 0 0nmol L反义寡核苷酸转染HeLa细胞 4 8h后 ,有 4条反义寡核苷酸对细胞增殖的抑制率超过 4 0 %,其中 4 5号反义寡核苷酸的抑制率可达5 9%,而阳性对照序列ISIS2 372 2的抑制率仅达 30 %.Northern和Western印迹分析证明 :4 5号反义寡核苷酸可明显降低细胞中survivin基因的mRNA含量和蛋白水平 .4 5号反义寡核苷酸还可在较低浓度 (2 0 0nmol L)显著增强HeLa细胞对化疗药三尖杉酯碱的敏感性 .因此 ,4 5号反义寡核苷酸有望应用于survivin高表达肿瘤的辅助治疗之中

Antisense Oligonucleotide Targeting Survivin Inhibits the Proliferation of Cancer Cells

Survivin is a novel gene encoding a structurally unique apoptosis inhibitor IAP. Abundantly expressed during fetal development, survivin is undetectable in terminally differentiated adult tissues. However, survivin is prominently expressed in transformed cell lines and in all the most common human cancers in vivo . To compare the abilities of inhibiting survivin expression, 20 human survivin antisense oligonucleotides were designed and selected by MTT assay. There are four human survivin antisense oligonucleotides having inhibitory rates above 40%, much higher than the inhibitory rate(about 30%) of the positive control(ISIS23722). The highest inhibitory rate (about 59%) was obtained by transfecting HeLa cells with antisense oligonucleotide No.45 for 24 hours. This antisense oligonucleotide's function of decreasing survivin expression was further demonstrated by Northern and Western blot. The experiment results demonstrate that antisense oligonucleotide No.45 can reduce survivin mRNA and protein level obviously. On the contrary, the mismatched control of the No.45 can not affect survivin expression. Furthermore, transfecting HeLa cells with the No.45 of 200 nmol/L can efficiently enhance cells' sensitivity to harringtonine. As the human survivin antisense oligonucleotide No.45 can decrease the expression of survivin in vitro and sensitize HeLa cells to chemotherapy, it deserves further investigation as a novel approach to selective tumor therapy.