Post-translational deregulation of YAP1 is genetically controlled in rat liver cancer and determines the fate and stem-like behavior of the human disease.
Post-translational deregulation of YAP1 is genetically controlled in rat liver cancer and determines the fate and stem-like behavior of the human disease.
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DOI：
10.18632/oncotarget.10246
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发表时间：
2016-08-02
期刊：
影响因子：
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通讯作者：
Pascale RM
中科院分区：
文献类型：
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作者：
Simile MM;Latte G;Demartis MI;Brozzetti S;Calvisi DF;Porcu A;Feo CF;Seddaiu MA;Daino L;Berasain C;Tomasi ML;Avila MA;Feo F;Pascale RM
先前的研究表明，YAP1 在肝细胞癌 (HCC) 中过度表达。在这里，我们观察到在遗传上易患肝癌的 F344 大鼠的发育不良结节和化学诱导的 HCC 中，Yap1/Ctgf 轴的表达高于在耐药 BN 大鼠中诱导的病变。在 BN 大鼠中，Yap1-tyr357、p73 磷酸化和 Caspase 3 裂解出现最高增加。在预后较差的人类 HCC 中（部分肝切除后生存期 < 3 年，HCCP），YAP1、CTGF、14-3-3 和 TEAD 蛋白以及 YAP1-14-3-3 和 YAP1-TEAD 复合物的水平高于预后较好的 HCC（生存期 > 3 年；HCCB）。在后者中，发生了更高水平的磷酸化 YAP1-ser127、YAP1-tyr357 和 p73、YAP1 泛素化和 Caspase 3 裂解。干性标记 NANOG、OCT-3/4 和 CD133 的表达在 HCCP 中最高，并且与 YAP1 和 YAP1-TEAD 水平相关。在 HepG2、Huh7 和 Hep3B 细胞中，强制 YAP1 过度表达会导致干细胞标记物表达并增加细胞活力，而通过特异性 siRNA 抑制 YAP1 表达或转染不与 TEAD 结合的突变体 YAP1，会诱导相反的变化。在用 YAP1 或 YAP1 siRNA 转染的 Huh7 细胞中，这些变化分别与细胞迁移和侵袭的刺激或抑制相关。此外，转录组分析表明，Huh7 细胞中 YAP1 转染可诱导与肿瘤干性相关的基因过度表达。总之，有利于 YAP1 泛素化和凋亡的翻译后修饰使 HCC 具有更好的预后，而有利于 YAP1-TEAD 复合物形成的条件与 HCC 细胞的侵袭性和干性特征的获得有关。
Previous studies showed that YAP1 is over-expressed in hepatocellular carcinoma (HCC). Here we observed higher expression of Yap1/Ctgf axis in dysplastic nodules and HCC chemically-induced in F344 rats, genetically susceptible to hepatocarcinogenesis, than in lesions induced in resistant BN rats. In BN rats, highest increase in Yap1-tyr357, p73 phosphorylation and Caspase 3 cleavage occurred. In human HCCs with poorer prognosis (< 3 years survival after partial liver resection, HCCP), levels of YAP1, CTGF, 14–3–3, and TEAD proteins, and YAP1-14-3-3 and YAP1-TEAD complexes were higher than in HCCs with better outcome (> 3 years survival; HCCB). In the latter, higher levels of phosphorylated YAP1-ser127, YAP1-tyr357 and p73, YAP1 ubiquitination, and Caspase 3 cleavage occurred. Expression of stemness markers NANOG, OCT-3/4, and CD133 were highest in HCCP and correlated with YAP1 and YAP1-TEAD levels. In HepG2, Huh7, and Hep3B cells, forced YAP1 over-expression led to stem cell markers expression and increased cell viability, whereas inhibition of YAP1 expression by specific siRNA, or transfection of mutant YAP1 which does not bind to TEAD, induced opposite alterations. These changes were associated, in Huh7 cells transfected with YAP1 or YAP1 siRNA, with stimulation or inhibition of cell migration and invasivity, respectively. Furthermore, transcriptome analysis showed that YAP1 transfection in Huh7 cells induces over-expression of genes involved in tumor stemness. In conclusion, Yap1 post-translational modifications favoring its ubiquitination and apoptosis characterize HCC with better prognosis, whereas conditions favoring the formation of YAP1-TEAD complexes are associated with aggressiveness and acquisition of stemness features by HCC cells.