Borealin-Derived Peptides as Survivin-Targeting Cancer Imaging and Therapeutic Agents
Borealin-Derived Peptides as Survivin-Targeting Cancer Imaging and Therapeutic Agents
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DOI：
10.1021/acs.bioconjchem.2c00398
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发表时间：
2022-11-01
影响因子：
4.7
通讯作者：
Fuchigami, Takeshi
中科院分区：
文献类型：
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作者：
Nozaki, Iori;Ishikawa, Natsumi;Fuchigami, Takeshi
Survivin在大多数癌细胞中过表达，但在正常成人组织中很少表达。它与预后差和对放射治疗和化学治疗的抵抗有关。在这项研究中，我们设计并合成了borealin衍生的小肽（Bor肽），作为生存素靶向药物用于癌症的诊断和治疗。这些肽对重组人存活素表现出结合亲和力（Kd = 49.6-193 nM），其中Bor 65 -75显示最高亲和力（Kd = 49.6 nM）。异硫氰酸荧光素标记的Bor 65 -75的荧光图像显示其与存活素在人胰腺癌细胞系MIA PaCa-2中的表达共定位。在WST-1测定中，细胞可穿透的九-D-精氨酸缀合的Bor 65 -75（r9-Bor 65 -75）抑制MIA PaCa-2细胞和MDA-MB-231细胞的生长（在10 μ M时分别抑制89和88%），而它对人乳腺上皮细胞系MCF-10A几乎没有影响，MCF-10A固有地不具有高存活素表达。Annexin V和碘化丙啶染色的流式细胞术显示，r9-Bor 65 -75以剂量依赖性方式诱导MIA PaCa 2细胞凋亡。通过蛋白质印迹法在暴露于r9-Bor 65 -75的MIA PaCa-2细胞中观察到裂解的聚ADP-核糖聚合酶蛋白表达的增加，表明r9-Bor 65 -75通过诱导凋亡抑制细胞增殖。在体内，r9-Bor 65 -75显著抑制MIA PaCa-2异种移植小鼠中的肿瘤生长，而没有任何显著的体重减轻。因此，Bor肽是用于开发以生存素为靶点的癌症成像和抗癌剂的有希望的候选物。
Survivin is overexpressed in most cancer cells but is rarely expressed in normal adult tissues. It is associated with poor prognosis and resistance to radiation therapy and chemotherapy. In this study, we designed and synthesized borealin-derived small peptides (Bor peptides) to function as survivin-targeting agents for the diagnosis and treatment of cancers. These peptides exhibited binding affinities for recombinant human survivin (Kd = 49.6-193 nM), with Bor65-75 showing the highest affinity (Kd = 49.6 nM). Fluorescence images of fluorescein isothiocyanate-labeled Bor65-75 showed its co-localization with survivin expression in the human pancreatic cancer cell line, MIA PaCa-2. In the WST-1 assay, cell penetrable nona-D-arginine-conjugated Bor65-75 (r9-Bor65-75) inhibited the growth of MIA PaCa-2 cells and MDA-MB-231 cells (89 and 88% inhibition at 10 mu M, respectively), whereas it had almost no effect on the human mammary epithelial cell line, MCF-10A, that inherently does not have high survivin expression. Flow cytometry with annexin V and propidium iodide staining revealed that r9-Bor65-75 induced apoptosis in MIA PaCa2 cells in a dose-dependent manner. An increase in cleaved poly ADP-ribose polymerase protein expression was observed in MIA PaCa-2 cells exposed to r9-Bor65-75 by western blotting, suggesting that r9-Bor65-75 inhibits cell proliferation by inducing apoptosis. In vivo, r9-Bor65-75 significantly suppressed tumor growth in MIA PaCa-2 xenograft mice, without any marked weight loss. Hence, Bor peptides are promising candidates for the development of cancer imaging and anticancer agents targeting survivin.