Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation.
Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation.
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Ajuba 通过靶向 β-连环蛋白和 YAP 信号传导抑制肝细胞癌细胞生长，并通过 neddylation 受 E3 连接酶 Hakai 调节
DOI：
10.1186/s13046-018-0806-3
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发表时间：
2018-07-24
期刊：
影响因子：
--
通讯作者：
Piao H
中科院分区：
文献类型：
--
作者：
Liu M;Jiang K;Lin G;Liu P;Yan Y;Ye T;Yao G;Barr MP;Liang D;Wang Y;Gong P;Meng S;Piao H
β-catenin和Yes相关蛋白（雅普）信号通路的异常激活与肝细胞癌（HCC）的进展相关。LIM结构域蛋白Ajuba调节β-连环蛋白和雅普信号传导，并与肿瘤发生有关。然而，Ajuba在HCC细胞中的表达的作用和机制仍不清楚。E3连接酶Hakai已被证明与其他Ajuba家族成员相互作用，Hakai是否相互作用并调节Ajuba尚不清楚。使用表达针对Ajuba或Hakai的shRNA的慢病毒建立稳定耗尽Ajuba或Hakai的HCC细胞系。通过体外贴壁生长、三维培养、跨孔侵袭实验等细胞学分析，研究了筋骨草对肝癌细胞的作用。在异种移植模型中测定体内肿瘤生长，并通过免疫组织化学检查肿瘤切片中的筋骨草表达。采用免疫共沉淀、共聚焦显微镜和免疫印迹法研究了筋骨草和哈卡伊之间的表达和相互作用。在异种移植模型中，HCC细胞中的筋骨草消耗显著增强了锚定非依赖性生长、侵袭、球体的形成和肿瘤生长，表明筋骨草在HCC中具有肿瘤抑制功能。从机制上讲，阿朱巴耗尽引发E-钙粘蛋白丢失和β-连环蛋白易位，细胞周期蛋白D1水平增加。此外，筋骨草的耗竭上调了雅普及其靶基因CYR 61的水平。此外，siRNA介导的β-连环蛋白或雅普的敲低减弱了通过消除Ajuba对HCC细胞的促肿瘤作用。值得注意的是，HCC细胞中的Ajuba稳定性受Hakai调节，Hakai是E-钙粘蛋白的E3连接酶。Hakai通过其HYB结构域与筋骨草相互作用，并诱导筋骨草neddylation，这被neddylation抑制剂MLN 4924拮抗，但不被MG 132拮抗。我们进一步表明，Hakai在HCC细胞中的过度表达显著增加了异种移植物中的锚定非依赖性生长、球状体形成能力和肿瘤生长，而Hakai耗竭导致了这些相反的效果，表明Hakai在HCC中的致癌作用。Hakai还诱导β-连环蛋白易位，增加细胞周期蛋白D1的水平。我们的数据表明，在HCC中的作用筋骨草和Hakai，并揭示机制的调节筋骨草的稳定性。本文的在线版本（10.1186/s13046-018-0806-3）包含补充材料，可供授权用户使用。
Aberrant activation of β-catenin and Yes-associated protein (YAP) signaling pathways has been associated with hepatocellular carcinoma (HCC) progression. The LIM domain protein Ajuba regulates β-catenin and YAP signaling and is implicated in tumorigenesis. However, roles and mechanism of Ajuba expression in HCC cells remain unclear. The E3 ligase Hakai has been shown to interact with other Ajuba family members and whether Hakai interacts and regulates Ajuba is unknown. HCC cell lines stably depleted of Ajuba or Hakai were established using lentiviruses expressing shRNAs against Ajuba or Hakai. The effects of Ajuba on HCC cells were determined by a number of cell-based analyses including anchorage-independent growth, three dimension cultures and trans-well invasion assay. In vivo tumor growth was determined in a xenograft model and Ajuba expression in tumor sections was examined by immunohistochemistry. Co-immunoprecipitation, confocal microscopy and immunoblot assay were used to examine the expression and interaction between Ajuba and Hakai. Depletion of Ajuba in HCC cells significantly enhanced anchorage-independent growth, invasion, the formation of spheroids and tumor growth in a xenograft model, suggesting a tumor suppressor function for Ajuba in HCC. Mechanistically, Ajuba depletion triggered E-cadherin loss and β-catenin translocation with increased Cyclin D1 levels. In addition, depletion of Ajuba upregulated the levels of YAP and its target gene CYR61. Furthermore, siRNA-mediated knockdown of either β-catenin or YAP attenuated the pro-tumor effects by Ajuba depletion on HCC cells. Notably, Ajuba stability in HCC cells was regulated by Hakai, an E3 ligase for E-cadherin. Hakai interacted with Ajuba via its HYB domain and induced Ajuba neddylation, which was antagonized by the neddylation inhibitor, MLN4924, but not MG132. We further show that overexpression of Hakai in HCC cells markedly increased anchorage-independent growth, spheroid-formation ability and tumor growth in xenografts whereas Hakai depletion resulted in these opposite effects, indicating an oncogenic role for Hakai in HCC. Hakai also induced β-catenin translocation with increased levels of Cyclin D1. Our data suggest a role for Ajuba and Hakai in HCC, and uncover the mechanism underlying the regulation of Ajuba stability. The online version of this article (10.1186/s13046-018-0806-3) contains supplementary material, which is available to authorized users.
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影响因子：
5.3
作者：
Jagannathan R;Schimizzi GV;Zhang K;Loza AJ;Yabuta N;Nojima H;Longmore GD
通讯作者：
Longmore GD
影响因子：
3.7
作者：
Abella V;Valladares M;Rodriguez T;Haz M;Blanco M;Tarrío N;Iglesias P;Aparicio LA;Figueroa A
通讯作者：
Figueroa A
影响因子：
11.8
作者：
Langer, Ellen M.;Feng, Yunfeng;Longmore, Gregory D.
通讯作者：
Longmore, Gregory D.
影响因子：
4.2
作者：
Hoshida Y;Toffanin S;Lachenmayer A;Villanueva A;Minguez B;Llovet JM
通讯作者：
Llovet JM
DOI：
10.1016/j.cub.2010.02.035
发表时间：
2010-04-13
期刊：
Current biology : CB
影响因子：
--
作者：
Das Thakur M;Feng Y;Jagannathan R;Seppa MJ;Skeath JB;Longmore GD
通讯作者：
Longmore GD