Silencing of survivin gene by small interfering RNAs produces supra-additive growth suppression in combination with 17-allylamino-17-demethoxygeldanamycin in human prostate cancer cells
Silencing of survivin gene by small interfering RNAs produces supra-additive growth suppression in combination with 17-allylamino-17-demethoxygeldanamycin in human prostate cancer cells
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DOI：
10.1158/1535-7163.mct-05-0132
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发表时间：
2006-01-01
影响因子：
5.7
通讯作者：
Zaffaroni, N
中科院分区：
文献类型：
--
作者：
Paduano, F;Villa, R;Zaffaroni, N
Survivin是一种抗凋亡基因，在大多数人类肿瘤中过表达，参与有丝分裂检查点调控。最近的证据表明，热休克蛋白90(Hsp90)在Survivin功能调节中起着重要作用。虽然Survivin-Hsp90的相关性可能促进肿瘤细胞的增殖，但它也可能为设计新的抗癌方法提供新的机会。我们评价了小干扰RNA(SiRNA)介导的Survivin抑制对前列腺癌细胞增殖能力的影响及其对Hsp90抑制剂17-烯丙氨基-17-去甲氧基格尔达那霉素(17-AAG)的敏感性。将4个针对Survivin基因不同部分的21聚体双链siRNA(100nmol/L)导入雄激素非依赖性前列腺癌DU145和PC-3细胞。转染后，收集细胞并分析Survivin基因和蛋白的表达、细胞增殖率、细胞的凋亡能力以及对17-AAG的敏感性。转染siRNAs的前列腺癌细胞可诱导不同程度的Survivin mRNA表达抑制(与对照组相比，抑制程度为39-60%)，同时Survivin蛋白丰度降低38%-75%。3种siRNAs对Survivin表达的抑制作用最强，可显著抑制细胞增殖，提高细胞凋亡率，并伴随caspase-9活性的升高。用siRNA和17-AAG连续处理诱导所有细胞系中超相加的抗增殖作用，并增强caspase-9依赖的凋亡反应。这些发现表明，旨在干扰Survivin-Hsp90连接的联合策略可能为雄激素非依赖性前列腺癌的治疗提供新的方法。
Survivin is an antiapoptotic gene, which is overexpressed in most human tumors and involved in mitotic checkpoint control. Recent evidence points to an essential role for heat shock protein 90 (Hsp90) in survivin function regulation. Although the survivin-Hsp90 association may promote tumor cell proliferation, it may also suggest new opportunities for the design of novel anticancer approaches. We evaluated the effect of small interfering RNA (siRNA)-mediated inhibition of survivin on the proliferative potential of prostate cancer cells and their sensitivity to the Hsp90 inhibitor 17-allylamino-17-demethoxygeldanamycin (17-AAG). Human androgen-independent prostate cancer cell lines (DU145 and PC-3) were transfected with four 21-mer double-stranded siRNAs (100 nmol/L) directed against different portions of survivin mRNA. After transfection, cells were collected and analyzed for survivin mRNA and protein expression, cell proliferation rate, ability to undergo apoptosis, and sensitivity to 17-AAG. Transfection of prostate cancer cells with siRNAs induced a variable extent of inhibition of survivin mRNA expression (39-60% compared with controls), which was paralleled by a 38% to 75% reduction in survivin protein abundance. The three siRNAs able to induce the greatest inhibition of survivin expression also significantly reduced cell proliferation and enhanced the rate of apoptosis, with a concomitant increase in caspase-9 activity. Sequential treatment with siRNA and 17-AAG induced supra-additive antiproliferative effects in all cell lines, with an enhanced caspase-9-dependent apoptotic response. These findings suggest that combined strategies aimed at interfering with the survivin-Hsp90 connection may provide novel approaches for treatment of androgen-independent prostate cancer.