Yhhu3813 is a novel selective inhibitor of c-Met kinase that inhibits c-Met-dependent neoplastic phenotypes of human cancer cells
Yhhu3813 is a novel selective inhibitor of c-Met kinase that inhibits c-Met-dependent neoplastic phenotypes of human cancer cells
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DOI：
10.1038/aps.2013.125
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发表时间：
2014-01-01
影响因子：
8.2
通讯作者：
Geng, Mei-yu
中科院分区：
文献类型：
--
作者：
He, Chang-xi;Ai, Jing;Geng, Mei-yu
目的：c-Met激酶失控与人类癌症的形成、发展和扩散密切相关。在本研究中，我们确定Yhhu3813为c-Met激酶的小分子抑制剂，并对其体内外抗肿瘤活性进行了研究。方法：采用ELISA法检测不同类型激酶的活性，用Western blotting检测细胞内信号蛋白的表达。用SRB法和四甲基偶氮唑盐比色法检测细胞增殖，用流式细胞仪检测细胞周期分布。Transwell法检测细胞的迁移和侵袭能力。形态发生实验检测细胞侵袭性生长。结果：Yhhu3813在体外能有效抑制c-Met的活性，其IC50值为2.4+/-0.3nmol/L，是15种不同酪氨酸激酶的400倍，表明Yhhu3813具有较高的选择性。该化合物(20,100和500nmol/L)剂量依赖性地抑制c-Met的磷酸化及其关键下游Akt和Erk信号级联在多种c-Met异常的人类癌细胞系中，无论c-Met激活跨不同细胞环境的机制的复杂性。在具有不同c-Met表达/激活背景的20株人癌细胞中，Yhhu3813通过将细胞阻滞在G(1)/S期而有效地抑制了c-Met诱导的细胞增殖。此外，Yhhu3813显著削弱了c-Met介导的细胞迁移、侵袭、分散和侵袭性生长。EBC-1移植瘤小鼠口服Yhhu3813(50或100 mg·kg(-1)·d(-1)，qd，连续2周)可剂量依赖性地抑制肿瘤生长，其抑制作用与肿瘤细胞增殖指数和c-Met信号转导有关。结论：Yhhu3813是一种有效的c-Met选择性抑制剂，在体内外均能抑制c-Met依赖的肿瘤表型。
Aim: c-Met kinase deregulation is strongly associated with the formation, progression and dissemination of human cancers. In this study we identified Yhhu3813 as a small-molecule inhibitor of c-Met kinase and characterized its antitumor properties both in vitro and in vivo.Methods: The activities of different kinases were measured using ELISA assays and signaling proteins in the cells were detected with Western blotting. Cell proliferation was assessed using SRB or MTT assay in twenty human cell lines and cell cycle distribution was determined with flow cytometry. Transwell-based assay was used to evaluate cell migration and invasion. Cell invasive growth was detected by a morphogenesis assay. c-Met overactivated human NSCLC cell line EBC-1 xenografts were used to evaluate the in vivo anti-tumor efficacy.Results: Yhhu3813 potently inhibited c-Met kinase activity in vitro with an IC50 value of 2.4 +/- 0.3 nmol/L, > 400-fold higher than that for a panel of 15 different tyrosine kinases, suggesting a high selectivity of Yhhu3813. The compound (20, 100 and 500 nmol/L) dose-dependently inhibited the phosphorylation of c-Met and its key downstream Akt and Erk signal cascades in multiple c-Met aberrant human cancer cell lines, regardless of the mechanistic complexity in c-Met activation across different cellular contexts. In 20 human cancer cell lines harboring different backgrounds of c-Met expression/activation, Yhhu3813 potently inhibited c-Met-driven cell proliferation via arresting cells at G(1)/S phase. Furthermore, Yhhu3813 substantially impaired c-Met-mediated cell migration, invasion, scattering, and invasive growth. Oral administration of EBC-1 xenograft mice with Yhhu3813 (50 or 100 mg.kg(-1).d(-1), qd, for 2 weeks) dose-dependently suppressed the tumor growth, which was correlated with a reduction in the intratumoral proliferation index and c-Met signaling.Conclusion: Yhhu3813 is a potent selective inhibitor of c-Met that inhibits c-Met-dependent neoplastic phenotypes of human cancer cells in vitro and in vivo.