Phosphoproteome profiling provides insight into the mechanism of action for carvedilol-mediated cancer prevention
Phosphoproteome profiling provides insight into the mechanism of action for carvedilol-mediated cancer prevention
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DOI：
10.1002/mc.22820
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发表时间：
2018-08-01
影响因子：
4.6
通讯作者：
Huang, Ying
中科院分区：
文献类型：
--
作者：
Cleveland, Kristan H.;Yeung, Steven;Huang, Ying
最近的研究表明，受体阻滞剂卡维地洛可以预防皮肤癌的发生，但其机制尚不清楚。卡维地洛是基于促进β-抑制蛋白介导的过程如ERK磷酸化的能力而被鉴定为偏性激动剂的少数β-阻断剂之一。为了了解磷酸化蛋白质组学信号在卡维地洛的抗癌活性中的作用，使用含有超过30个信号传导途径的1318个位点特异性和磷酸化特异性抗体的Phospho Explorer抗体阵列分析用EGF、卡维地洛或其组合处理的小鼠表皮JB 6 P+细胞。阵列数据表明，EGF和卡维地洛都增加ERK的胞浆靶点P70 S6 K的磷酸化，而其核靶点ELK-1仅被EGF激活;此外，卡维地洛减弱EGF诱导的ELK-1和c-Jun的磷酸化。亚细胞分级分析表明，EGF诱导的ERK核转位被卡维地洛阻断。Western印迹和荧光素酶报告基因分析证实，偏向性阻断剂卡维地洛和阿普洛尔阻断EGF诱导的c-Jun/AP-1和ELK-1的磷酸化和活化。一致地，卡维地洛和阿普洛尔都强烈地阻止EGF诱导的JB 6 P+细胞的肿瘤转化。值得注意的是，口服卡维地洛治疗显著抑制了SCID小鼠中A375黑素瘤异种移植物的生长。由于ERK的核转位是癌发生的关键步骤，因此提出抑制该事件作为偏向性阻断剂如卡维地洛的新的抗癌机制。
Recent studies suggest that the -blocker drug carvedilol prevents skin carcinogenesis but the mechanism is unknown. Carvedilol is one of a few -blockers identified as biased agonist based on an ability to promote -arrestin-mediated processes such as ERK phosphorylation. To understand the role of phosphoproteomic signaling in carvedilol's anticancer activity, the mouse epidermal JB6 P+ cells treated with EGF, carvedilol, or their combination were analyzed using the Phospho Explorer Antibody Array containing 1318 site-specific and phospho-specific antibodies of over 30 signaling pathways. The array data indicated that both EGF and carvedilol increased phosphorylation of ERK's cytosolic target P70S6K while its nuclear target ELK-1 were activated only by EGF; Furthermore, EGF-induced phosphorylation of ELK-1 and c-Jun was attenuated by carvedilol. Subcellular fractionation analysis indicated that ERK nuclear translocation induced by EGF was blocked by co-treatment with carvedilol. Western blot and luciferase reporter assays confirmed that the biased -blockers carvedilol and alprenolol blocked EGF-induced phosphorylation and activation of c-Jun/AP-1 and ELK-1. Consistently, both carvedilol and alprenolol strongly prevented EGF-induced neoplastic transformation of JB6 P+ cells. Remarkably, oral carvedilol treatment significantly inhibited the growth of A375 melanoma xenograft in SCID mice. As nuclear translocation of ERK is a key step in carcinogenesis, inhibition of this event is proposed as a novel anticancer mechanism for biased -blockers such as carvedilol.