Glutamatergic pathway targeting in melanoma: single-agent and combinatorial therapies.
Glutamatergic pathway targeting in melanoma: single-agent and combinatorial therapies.
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DOI：
10.1158/1078-0432.ccr-11-0098
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发表时间：
2011-11-15
期刊：
影响因子：
--
通讯作者：
Chen S
中科院分区：
文献类型：
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作者：
Lee HJ;Wall BA;Wangari-Talbot J;Shin SS;Rosenberg S;Chan JL;Namkoong J;Goydos JS;Chen S
黑色素瘤是一种异质性疾病，其中单一疗法可能由于基因组特征的变化而失败。B-RAF抑制剂在临床上是不充分的，但通过靶向多个信号通路的联合治疗可能会增强反应。我们研究了多激酶抑制剂索拉非尼或突变的B-RAF抑制剂PLX 4720与阿曲唑（一种拮抗黑色素瘤细胞中GRM 1（代谢型谷氨酸受体1）信号传导的谷氨酸释放抑制剂）联合使用的临床前疗效。表达GRM 1和野生型B-RAF或突变型B-RAF的黑素瘤细胞系用阿鲁唑、索拉非尼、PLX 4720或阿鲁唑与索拉非尼或与PLX 4720的组合处理。细胞外谷氨酸水平通过谷氨酸释放测定来确定。MTT测定和细胞周期分析证明了化合物对增殖、活力和细胞周期谱的影响。Western免疫印迹和免疫组化染色显示凋亡标记。通过蛋白质免疫印迹评估MAPK途径的后果。异种移植肿瘤模型用于确定化合物的体内功效。阿鲁唑与索拉非尼的组合在携带野生型或突变的B-RAF的表达GRM 1的黑素瘤细胞中表现出增强的抗肿瘤活性。与阿舒唑和索拉非尼组合相比，阿舒唑与PLX 4720的组合在抑制具有B-RAFV 600 E突变的表达GRM 1的细胞的生长方面显示出降低的功效。无论B-RAF基因型如何，阿鲁唑与索拉非尼的组合在表达GRM 1的黑素瘤细胞中在体外和体内抑制肿瘤增殖方面似乎是有效的，并且可能是可行的治疗性临床组合。
Melanoma is a heterogeneous disease where monotherapies are likely to fail due to variations in genomic signatures. B-RAF inhibitors have been clinically inadequate but response might be augmented with combination therapies targeting multiple signaling pathways. We investigate the pre-clinical efficacy of combining the multi-kinase inhibitor Sorafenib or mutated B-RAF inhibitor PLX4720 with Riluzole, an inhibitor of glutamate release that antagonizes GRM1 (metabotropic glutamate receptor1) signaling in melanoma cells. Melanoma cell lines that express GRM1 and either wild type B-RAF or mutated B-RAF were treated with Riluzole, Sorafenib, PLX4720 or the combination of Riluzole with Sorafenib or with PLX4720. Extra-cellular glutamate levels were determined by glutamate release assays. MTT assays and cell cycle analysis demonstrate effects of the compounds on proliferation, viability and cell cycle profiles. Western immunoblots and immunohistochemical staining showed apoptotic markers. Consequences on MAPK pathway were assessed by western immunoblots. Xenograft tumor models were used to determine the efficacy of the compounds in vivo. The combination of Riluzole with Sorafenib exhibited enhanced anti-tumor activities in GRM1 expressing melanoma cells harboring either wild type or mutated B-RAF. The combination of Riluzole with PLX4720 showed lessened efficacy compared with the Riluzole and Sorafenib combination in suppressing the growth of GRM1 expressing cells harboring the B-RAFV600E mutation. The combination of Riluzole with Sorafenib appears potent in suppressing tumor proliferation in vitro and in vivo in GRM1 expressing melanoma cells regardless of B-RAF genotype and may be a viable therapeutic clinical combination.