Impairment of glioma stem cell survival and growth by a novel inhibitor for Survivin-Ran protein complex.
Impairment of glioma stem cell survival and growth by a novel inhibitor for Survivin-Ran protein complex.
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DOI：
10.1158/1078-0432.ccr-12-0647
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发表时间：
2013-02-01
期刊：
影响因子：
--
通讯作者：
Nakano I
中科院分区：
文献类型：
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作者：
Guvenc H;Pavlyukov MS;Joshi K;Kurt H;Banasavadi-Siddegowda YK;Mao P;Hong C;Yamada R;Kwon CH;Bhasin D;Chettiar S;Kitange G;Park IH;Sarkaria JN;Li C;Shakhparonov MI;Nakano I
多形性胶质母细胞瘤（GBM）是一种毁灭性的疾病。最近的研究表明，GBM的干细胞特性有助于治疗抗性的发展。Survivin和Ran的表达通过GBM组织的免疫组织化学、定量逆转录酶（qRT）-PCR和患者来源的GBM球培养物的免疫细胞化学来评估。利用基于计算结构的药物设计，设计、合成了11个小分子化合物，并评估了它们作为Survivin蛋白分子相互作用的抑制剂候选物。通过Western blot、ELISA、原位邻位连接试验和免疫细胞化学等方法研究了先导化合物LLP-3的作用机制。在体外和体内评估LLP-3处理对GSC的影响。采用免疫组化方法检测44例初诊和31例放化疗后复发GBM患者组织中Survivin的表达。最后，在体外评价了替莫唑胺耐药GBM球对LLP-3的敏感性。Survivin和Ran在GBM组织中强烈表达，特别是在血管周围，也在患者来源的GSC培养物中。LLP-3处理破坏了癌细胞中的Survivin-Ran蛋白复合物，并在体外和体内消除了患者来源的GBM球体的生长。这种抑制依赖于caspase活性，并与细胞的p53状态有关。免疫组化结果显示，复发性GBM中Survivin的表达显著高于新诊断的肿瘤，且替莫唑胺耐药GBM球对LLP-3治疗具有高敏感性。通过LLP-3破坏存活素-Ran复合物在体外和体内都消除了GSC的存活和生长，表明GBM的有吸引力的新治疗方法。
Glioblastoma multiforme (GBM) is a devastating disease. Recent studies suggest that the stem cell properties of GBM contribute to the development of therapy resistance. The expression of Survivin and Ran was evaluated by immunohistochemistry with GBM tissues, and quantitative reverse transcriptase (qRT)-PCR and immunocytochemistry with patient-derived GBM sphere cultures. With a computational structure-based drug design, 11 small-molecule compounds were designed, synthesized, and evaluated as inhibitor candidates for the molecular interaction of Survivin protein. The molecular mechanism of the lead compound, LLP-3, was determined by Western blot, ELISA, in situ proximity ligation assay, and immunocytochemistry. The effects of LLP-3 treatment on GSCs were evaluated both in vitro and in vivo. Quantitative immunohistochemistry was carried out to compare Survivin expression in tissues from 44 newly diagnosed and 31 recurrent post-chemoradiation GBM patients. Lastly, the sensitivities of temozolomide-resistant GBM spheres to LLP-3 were evaluated in vitro. Survivin and Ran were strongly expressed in GBM tissues, particularly in the perivasculature, and also in patient-derived GSC cultures. LLP-3 treatment disrupted the Survivin–Ran protein complex in cancer cells and abolished the growth of patient-derived GBM spheres in vitro and in vivo. This inhibition was dependent on caspase activity and associated with p53 status of cells. Immunohistochemistry showed that Survivin expression is significantly increased in recurrent GBM compared with newly diagnosed tumors, and temozolomide-resistant GBM spheres exhibited high sensitivities to LLP-3 treatment. Disruption of the Survivin–Ran complex by LLP-3 abolishes survival and growth of GSCs both in vitro and in vivo, indicating an attractive novel therapeutic approach for GBM.