Silencing of bcr/abl Chimeric Gene in Human Chronic Myelogenous Leukemia Cell Line K562 by siRNA-Nuclear Export Signal Peptide Conjugates.
Silencing of bcr/abl Chimeric Gene in Human Chronic Myelogenous Leukemia Cell Line K562 by siRNA-Nuclear Export Signal Peptide Conjugates.
复制标题
通过 siRNA-核输出信号肽缀合物沉默人慢性粒细胞白血病细胞系 K562 中的 bcr/abl 嵌合基因。
DOI：
10.1089/nat.2016.0647
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发表时间：
2017
影响因子：
4
通讯作者：
Fujii M.
中科院分区：
文献类型：
--
作者：
Shinkai S;Kashihara S;Minematsu G;Fujii H;Naemura M;Kotake Y;Morita Y;Ohnuki K;Fokina AA;Stetsenko DA;Filichev VV;Fujii M.
本文报道了用固相片段偶联法合成siRNA-NES（nuclear export signal）多肽偶联物，并将其应用于人慢性粒细胞白血病细胞系K562中bcr/abl嵌合基因的沉默。制备了两种类型的siRNA-内斯缀合物，其5′端的两条正义链分别与来自TFIIIA和HIV-1 REV的内斯肽共价连接。观察到它们的沉默效率显著增强。siRNA-TFIIIA内斯偶联物在200 nM和50 nM时对BCR/ABL基因的表达抑制率分别为8.3%和11.6%，siRNA-HIV-1 REV内斯偶联物在200 nM和50 nM时对BCR/ABL基因的表达抑制率分别为4.0%和6.3%，而天然siRNA在200 nM和50 nM时对BCR/ABL基因的表达抑制率分别为36.3%和30.2%。我们还可以显示siRNA-NES缀合物与所设计的两亲性肽肽β 7的复合物可以被细胞摄取而没有细胞毒性，并且显示出优异的沉默效率。我们相信，复合物siRNA-NES缀合物和肽β 7是一个有希望的候选人在体内和治疗应用。
Herein we described the synthesis of siRNA-NES (nuclear export signal) peptide conjugates by solid phase fragment coupling and the application of them to silencing of bcr/abl chimeric gene in human chronic myelogenous leukemia cell line K562. Two types of siRNA-NES conjugates were prepared, and both sense strands at 5′ ends were covalently linked to a NES peptide derived from TFIIIA and HIV-1 REV, respectively. Significant enhancement of silencing efficiency was observed for both of them. siRNA-TFIIIA NES conjugate suppressed the expression ofBCR/ABLgene to 8.3% at 200 nM and 11.6% at 50 nM, and siRNA-HIV-1REV NES conjugate suppressed to 4.0% at 200 nM and 6.3% at 50 nM, whereas native siRNA suppressed to 36.3% at 200 nM and 30.2% at 50 nM. We could also show complex of siRNA-NES conjugate and designed amphiphilic peptidepeptideβ7could be taken up into cells with no cytotoxicity and showed excellent silencing efficiency. We believe that the complex siRNA-NES conjugate andpeptideβ7is a promising candidate forin vivouse and therapeutic applications.