Protein kinase C inhibitor AEB071 targets ocular melanoma harboring GNAQ mutations via effects on the PKC/Erk1/2 and PKC/NF-κB pathways.
Protein kinase C inhibitor AEB071 targets ocular melanoma harboring GNAQ mutations via effects on the PKC/Erk1/2 and PKC/NF-κB pathways.
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DOI：
10.1158/1535-7163.mct-12-0121
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发表时间：
2012-09
影响因子：
5.7
通讯作者：
Hodi FS
中科院分区：
文献类型：
--
作者：
Wu X;Li J;Zhu M;Fletcher JA;Hodi FS
在大约50%的葡萄膜黑色素瘤（UM）中发现了209位密码子的体细胞GNAQ突变，并已报道通过激活PLCβ-PKC-Erk 1/2途径致癌。我们假设PKC可能为治疗携带GNAQ突变的UM提供新的机会。为了验证这一假设，将携带野生型或突变型GNAQ的UM细胞用PKC抑制剂AEB 071（sotraflavin）处理或用表达靶向PKC亚型的shRNA的慢病毒感染。值得注意的是，AEB 071在低微摩尔浓度下通过诱导G1期阻滞和细胞凋亡显著抑制携带GNAQ突变的UM细胞的生长。然而，AEB 071对携带野生型GNAQ的UM细胞几乎没有影响。在GNAQ突变的UM中，AEB 071介导的细胞抑制伴随着Erk 1/2磷酸化、NF-κB的抑制，细胞周期蛋白D1、生存素、Bcl-xL和XIAP的表达降低，以及细胞周期蛋白依赖性激酶抑制剂p27 Kip 1的表达增加。AEB 071抑制GNAQ突变UM细胞PKC α、β、δ、ε和θ的表达。我们的研究结果从shRNA介导的敲除研究表明，这些PKC亚型是功能上的重要UM细胞窝藏GNAQ突变。此外，Erk 1/2和NF-κB途径的抑制剂降低UM细胞的活力。总之，我们的研究结果表明，AEB 071通过靶向PKC/Erk 1/2和PKC/NF-κB通路对携带GNAQ突变的UM细胞发挥抗肿瘤作用。靶向PKC抑制药物如AEB 071为携带GNAQ突变的UM提供了新的治疗潜力。
Somatic GNAQ mutations at codon 209 have been identified in approximately 50% of uveal melanomas (UM) and have been reported to be oncogenic through activating PLCβ-PKC-Erk1/2 pathways. We hypothesized that PKC may provide new opportunities for therapeutic targeting of UM carrying GNAQ mutations. To test this hypothesis, UM cells harboring wild type or mutant GNAQ were treated with the PKC inhibitor AEB071 (sotrastaurin) or infected with lentivirus expressing shRNAs targeting PKC isoforms. Notably, AEB071 at low micromolar concentrations significantly inhibited the growth of UM cells harboring GNAQ mutations through induction of G1 arrest and apoptosis. However, AEB071 had little effect on UM cells carrying wild type GNAQ. AEB071-mediated cell inhibition in the GNAQ mutated UM was accompanied by inhibition of Erk1/2 phosphorylation, NF-κB, decreased expression of cyclin D1, survivin, Bcl-xL and XIAP, and increased expression of cyclin-dependent kinase inhibitor p27Kip1. AEB071 suppressed the expression of PKC α, β, δ, ε and θ in GNAQ mutated UM cells. Our findings from shRNA-mediated knockdown studies revealed that these PKC isoforms are functionally important for UM cells harboring GNAQ mutations. Furthermore, inhibitors of Erk1/2 and NF-κB pathways reduced viability of UM cells. Together, our findings demonstrate that AEB071 exerts antitumor action on UM cells carrying GNAQ mutations via targeting PKC/Erk1/2 and PKC/NF-κB pathways. Targeted PKC inhibition with drugs such as AEB071 offers novel therapeutic potential for UM harboring GNAQ mutations.