Small interfering RNA targeting survivin sensitizes lung cancer cell with mutant p53 to adriamycin
Small interfering RNA targeting survivin sensitizes lung cancer cell with mutant p53 to adriamycin
复制标题
DOI：
10.1002/ijc.21350
复制
发表时间：
2006-02-15
影响因子：
6.4
通讯作者：
Nakagawa, K
中科院分区：
文献类型：
--
作者：
Yonesaka, K;Tamura, K;Nakagawa, K
Survivin是凋亡抑制蛋白（IAP）家族的成员，其在癌组织中特异性过表达。p53是肿瘤抑制基因之一;其响应于DNA损伤的诱导引起细胞凋亡并与药物敏感性相关。为了研究p53对生存素的可能调控，我们检测了肺癌细胞系中生存素对阿霉素的反应水平。野生型p53的细胞系中生存素的mRNA和蛋白水平在p53诱导后显著下降，但在突变或无效的p53的细胞系中没有观察到生存素的这种减少。小干扰RNA对A549细胞中野生型p53的抑制作用显著上调survivin的表达。在p53突变的PC 9细胞中通过siRNA抑制Survivin显著抑制细胞增殖。为了研究抑制survivin后癌细胞对阿霉素的敏感性，我们使用siRNA抑制survivin表达，然后以IC 50剂量加入阿霉素。与阿霉素进一步孵育48小时后，与靶向扰乱的siRNA相比，用靶向生存素的siRNA处理的细胞的增殖显着抑制。此外，TUNEL和pro-caspase 3表达检测均显示阿霉素和靶向Survivin的siRNA联合处理后细胞凋亡显著增加。我们的研究结果表明，生存素下调p53，和siRNA靶向生存素增加细胞对阿霉素的敏感性，促进凋亡。靶向生存素的siRNA可能用于增加对抗癌药物的敏感性，特别是在具有突变的p53的耐药细胞中。(c)2005 Wiley-Liss，Inc.
Survivin is a member of the inhibitor of apoptosis protein (IAP) family that is specifically overexpressed in cancer tissues. p53 is one of the tumor suppressor genes; its induction in response to DNA damage causes apoptosis and correlates with drug sensitivity. To investigate the possible regulation of survivin by p53, we examined the level of survivin expression in lung cancer cell lines in response to adriamycin. Levels of survivin mRNA and protein in cell lines with wild-type p53 decreased dramatically after p53 induction, but no such reduction of survivin was observed in cell lines with mutated or null p53. Inhibition of wild-type p53 in A549 cells by small interfering (si) RNA. significantly upregulated the expression of survivin. Survivin inhibition by siRNA in PC9 cells with mutated p53 significantly depressed cell proliferation. To investigate the sensitivity of cancer cells to adriamycin after inhibition of survivin, we depressed survivin expression using siRNA, and then added adriamycin at an IC50 dose. After a further 48 hr incubation with adriamycin, proliferation was significantly depressed in the cells treated with siRNA targeting survivin, in comparison with siRNA targeting scramble. Furthermore, both TUNEL and pro-caspase3 expression assay showed a significant increase in apoptosis after combined treatment with adriamycin and siRNA targeting survivin. Our results demonstrate that survivin is downregulated by p53, and that siRNA targeting of survivin increases cell sensitivity to adriamycin and promotes apoptosis. siRNA targeting of survivin could be potentially useful for increasing sensitivity to anticancer drugs, especially in drug-resistant cells with mutated p53. (c) 2005 Wiley-Liss, Inc.