Aurora kinase targeting in lung cancer reduces KRAS-induced transformation.
Aurora kinase targeting in lung cancer reduces KRAS-induced transformation.
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DOI：
10.1186/s12943-016-0494-6
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发表时间：
2016-02-03
期刊：
影响因子：
37.3
通讯作者：
Bassères DS
中科院分区：
文献类型：
--
作者：
Dos Santos EO;Carneiro-Lobo TC;Aoki MN;Levantini E;Bassères DS
KRAS的激活突变在肺癌中普遍存在，并与肿瘤发生过程有因果联系。然而，针对致癌RAS的治疗到目前为止都是无效的，识别影响致癌表型的KRAS靶点是有必要的。根据已发表的研究表明，有丝分裂酶Aurora A(AURKA)和B(AURKB)与致癌的RAS协同促进恶性转化，并且AURKA使RAS效应通路组分磷酸化，本研究的目的是探讨AURKA和AURKB是否是肺癌的KRAS靶点，以及靶向这些激酶是否对治疗有益。为了确定致癌的KRAS是否诱导Aurora激酶的表达，我们在三种不同的基于肺细胞的KRAS功能获得或丧失模型中使用了qPCR和Western blotting。为了确定这些激酶在KRAS诱导的转化中的功能作用，我们建立了KRAS阳性的A549和H358细胞，通过稳定和可诱导的shRNA下调AURKA或AURKB，并评估了体外转化和体内肿瘤生长。为了验证AURKA和/或AURKB作为肺癌治疗相关的KRAS靶点，我们用双重Aurora激酶抑制剂处理A549和H358细胞，以及两种不同的基于肺细胞的KRAS功能获得模型，并进行了体外功能分析。我们确定KRAS正向调节AURKA和AURKB的表达。此外，在KRAS阳性的H358和A549细胞系中，AURKA或AURKB的可诱导基因敲除以及AURKA/AURKB双重抑制剂的处理，在体外会降低细胞的生长、活力、增殖、转化和诱导细胞凋亡。此外，可诱导的shRNA介导的A549细胞中AURKA基因的敲除降低了体内肿瘤的生长。更重要的是，AURKA和AURKB的双重药物抑制剂在体外以致癌的KRAS依赖的方式抑制生长、存活、转化和诱导细胞凋亡，表明Aurora激酶抑制疗法可以特异性地靶向KRAS转化的细胞。我们的结果支持我们的假设，即极光激酶是肺癌中KRAS的重要靶点，并提示极光激酶抑制是KRAS诱导的肺癌治疗的一种新方法。本文的在线版本(doi：10.1186/s12943-0160494-6)包含补充材料，授权用户可以使用。
Activating mutations in KRAS are prevalent in lung cancer and have been causally linked to the oncogenic process. However, therapies targeted to oncogenic RAS have been ineffective to date and identification of KRAS targets that impinge on the oncogenic phenotype is warranted. Based on published studies showing that mitotic kinases Aurora A (AURKA) and B (AURKB) cooperate with oncogenic RAS to promote malignant transformation and that AURKA phosphorylates RAS effector pathway components, the aim of this study was to investigate whether AURKA and AURKB are KRAS targets in lung cancer and whether targeting these kinases might be therapeutically beneficial. In order to determine whether oncogenic KRAS induces Aurora kinase expression, we used qPCR and western blotting in three different lung cell-based models of gain- or loss-of-function of KRAS. In order to determine the functional role of these kinases in KRAS-induced transformation, we generated KRAS-positive A549 and H358 cells with stable and inducible shRNA-mediated knockdown of AURKA or AURKB and evaluated transformation in vitro and tumor growth in vivo. In order to validate AURKA and/or AURKB as therapeutically relevant KRAS targets in lung cancer, we treated A549 and H358 cells, as well as two different lung cell based models of gain-of-function of KRAS with a dual Aurora kinase inhibitor and performed functional in vitro assays. We determined that KRAS positively regulates AURKA and AURKB expression. Furthermore, in KRAS-positive H358 and A549 cell lines, inducible knockdown of AURKA or AURKB, as well as treatment with a dual AURKA/AURKB inhibitor, decreased growth, viability, proliferation, transformation, and induced apoptosis in vitro. In addition, inducible shRNA-mediated knockdown of AURKA in A549 cells decreased tumor growth in vivo. More importantly, dual pharmacological inhibiton of AURKA and AURKB reduced growth, viability, transformation, and induced apoptosis in vitro in an oncogenic KRAS-dependent manner, indicating that Aurora kinase inhibition therapy can specifically target KRAS-transformed cells. Our results support our hypothesis that Aurora kinases are important KRAS targets in lung cancer and suggest Aurora kinase inhibition as a novel approach for KRAS-induced lung cancer therapy. The online version of this article (doi:10.1186/s12943-016-0494-6) contains supplementary material, which is available to authorized users.