Suppression of FOXM1 Transcriptional Activities via a Single-Stranded DNA Aptamer Generated by SELEX.
Suppression of FOXM1 Transcriptional Activities via a Single-Stranded DNA Aptamer Generated by SELEX.
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通过 SELEX 生成的单链 DNA 适体抑制 FOXM1 转录活性
DOI：
10.1038/srep45377
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发表时间：
2017-03-30
影响因子：
4.6
通讯作者：
Tan Y
中科院分区：
文献类型：
--
作者：
Xiang Q;Tan G;Jiang X;Wu K;Tan W;Tan Y
转录因子 FOXM1 通过其 DNA 结合域 (DBD) 与其启动子处的共有序列结合，并激活增殖相关基因。 FOXM1 的异常过度表达与许多癌症的发生和进展相关。抑制 FOXM1 转录活性被提议作为癌症治疗的潜在治疗策略。在本研究中，我们通过SELEX获得了FOXM1特异性单链DNA适体（FOXM1 Apt），并以重组FOXM1 DBD蛋白作为选择靶点。通过电泳迁移率变动测定 (EMSA) 和荧光偏振 (FP) 测定证实了 FOXM1 Apt 与 FOXM1 蛋白的结合。硫代磷酸酯修饰的 FOXM1 Apt (M-FOXM1 Apt) 作为野生型 FOXM1 Apt 与 FOXM1 结合，并与 FOXM1 共定位于细胞核中。 M-FOXM1-Apt 消除了 FOXM1 在其共有结合位点上的结合并抑制了 FOXM1 转录活性。与癌细胞中FOXM1的RNA干扰相比，M-FOXM1 Apt在不改变FOXM1水平的情况下抑制细胞增殖和FOXM1靶基因的表达。我们的结果表明，获得的FOXM1 Apt可以用作FOXM1检测的探针，同时可以用作癌细胞中FOXM1转录功能的抑制剂，为未来的癌症诊断和治疗提供潜在的试剂。
The transcription factor FOXM1 binds to its consensus sequence at promoters through its DNA binding domain (DBD) and activates proliferation-associated genes. The aberrant overexpression of FOXM1 correlates with tumorigenesis and progression of many cancers. Inhibiting FOXM1 transcriptional activities is proposed as a potential therapeutic strategy for cancer treatment. In this study, we obtained a FOXM1-specific single stranded DNA aptamer (FOXM1 Apt) by SELEX with a recombinant FOXM1 DBD protein as the target of selection. The binding of FOXM1 Apt to FOXM1 proteins were confirmed with electrophoretic mobility shift assays (EMSAs) and fluorescence polarization (FP) assays. Phosphorthioate-modified FOXM1 Apt (M-FOXM1 Apt) bound to FOXM1 as wild type FOXM1 Apt, and co-localized with FOXM1 in nucleus. M-FOXM1-Apt abolished the binding of FOXM1 on its consensus binding sites and suppressed FOXM1 transcriptional activities. Compared with the RNA interference of FOXM1 in cancer cells, M-FOXM1 Apt repressed cell proliferation and the expression of FOXM1 target genes without changing FOXM1 levels. Our results suggest that the obtained FOXM1 Apt could be used as a probe for FOXM1 detection and an inhibitor of FOXM1 transcriptional functions in cancer cells at the same time, providing a potential reagent for cancer diagnosis and treatment in the future.