Knockdown of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independently of p53
Knockdown of survivin expression by small interfering RNA reduces the clonogenic survival of human sarcoma cell lines independently of p53
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DOI：
10.1038/sj.cgt.7700677
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发表时间：
2004-03-01
影响因子：
6.4
通讯作者：
Taubert, H
中科院分区：
文献类型：
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作者：
Kappler, M;Bache, M;Taubert, H
Survivin是凋亡抑制基因家族的一员，在许多肿瘤类型中过度表达。Survivin是软组织肉瘤的预后标志物，但在这些肿瘤中Survivin表达下调以及Survivin下调对p53的依赖性尚未被研究。因此，我们应用小干扰RNA （siRNA）在5种具有野生型或突变型p53等位基因的人肉瘤细胞系中敲低survivin的表达。与无意义siRNA处理的肉瘤细胞系中survivin mRNA的表达相比，survivin特异性siRNA处理后的表达降低了73-88%；Survivin蛋白表达降低52-81%。这一发现与克隆存活率降低65% -86%相结合。然而，用survivin特异性siRNA处理的细胞中只有不到10%发生凋亡。细胞周期和形态学分析表明，在G2/M期处理细胞数量急剧增加后，部分细胞变成多倍体；这一结果表明，大量处理过的细胞有丝分裂是不完整的。我们的研究结果表明，无论是否存在野生型p53等位基因，survivin特异性siRNA都可能是一种选择性杀死肉瘤细胞的治疗方法。
Survivin, a member of the inhibitors-of-apoptosis gene family, is overexpressed in many tumor types. Survivin is a prognostic marker of soft-tissue sarcomas, but the downregulation of survivin expression and the possible dependency of survivin downregulation on p53 in these tumors have not been investigated. Therefore, we applied small interfering RNA (siRNA) to knock down the expression of survivin in five human sarcoma cell lines with wild-type or mutant p53 alleles. Compared with survivin mRNA expression in the nonsense siRNA-treated sarcoma cell lines, expression after treatment with survivin-specific siRNA was reduced by 73-88%; survivin protein expression was reduced by 52-81%. This finding was coupled with a reduction in clonogenic survival ranging from 65-86%. However, less than 10% of cells treated with survivin-specific siRNA underwent apoptosis. Cell-cycle and morphologic analyses showed that after a dramatic increase in the number of treated cells in the G2/M phase, some of the cells became polyploid; this result indicates that mitosis of a substantial number of treated cells was incomplete. Our findings suggest that survivin-specific siRNA could be a selective treatment to kill sarcoma cells regardless of the presence or absence of wild-type p53 alleles.