WWC3 inhibits epithelial-mesenchymal transition of lung cancer by activating Hippo-YAP signaling.
WWC3 inhibits epithelial-mesenchymal transition of lung cancer by activating Hippo-YAP signaling.
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WWC3通过激活Hippo-YAP信号抑制肺癌上皮间质转化
DOI：
10.2147/ott.s162387
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发表时间：
2018
影响因子：
4
通讯作者：
Wang E
中科院分区：
文献类型：
--
作者：
Han Q;Kremerskothen J;Lin X;Zhang X;Rong X;Zhang D;Wang E
虽然我们最近报道WWC3通过激活HIPPO通路来抑制肺癌的侵袭和转移，但这一过程的影响和潜在机制仍不清楚。为明确WWC3在肺癌上皮间充质转化中的作用，我们采用免疫组织化学方法检测了WWC3和EMT相关生物标记物在127例NSCLC患者中的表达水平，并分析了它们之间的相关性。采用创伤愈合实验和细胞侵袭实验检测WWC3基因敲除后细胞侵袭能力的变化。采用定量逆转录聚合酶链式反应(QRT-PCR)和免疫印迹法检测EMT相关生物标记物的mRNA和蛋白水平，以及WWC3引起的河马信号转导主要分子的变化。免疫沉淀法检测WWC3与LATS1的相互作用。WWC3基因敲除导致肺癌细胞从上皮型向间充质表型的显著转变。此外，WWC3在肺癌细胞中的异位表达降低了间充质标志物的表达，增加了上皮标志物的表达，但WWC3-ΔWW质粒取消了这一作用。ShRNA沉默WWC3则起到相反的作用。此外，WWC3水平与肺癌细胞和标本中EMT诱导物(Snail和Slug)的水平呈负相关。免疫印迹显示，WWC3野生型通过其WW结构域上调大肿瘤抑制因子(LATS1)和YAP的磷酸化，从而激活河马通路。敲除YAP和LATS1，以及维替普芬(VP)的使用，可以逆转WWC3沉默所引起的这一效应。这些发现表明，WWC3作为一种肿瘤抑制因子，抑制了EMT过程，使其成为肺癌潜在的治疗靶点。
Though we recently reported that the WWC3 inhibits the invasiveness and metastasis of lung cancer by activating the Hippo pathway, the impact and underlying mechanisms of this process still remain unclear. To identify the role of WWC3 in epithelial-mesenchymal transition of lung cancer, we performed immunohistochemistry to detect the expression levels of WWC3 and EMT-related biomarker, and analyzed their correlations in a cohort of 127 patients with NSCLC. Wound healing assay and cell invasion assay were applied to explore cell invasive ability change after WWC3 knockdown. qRT-PCR and immunoblotting were performed to assess mRNA and protein levels of EMT-related biomarkers and the main molecules changes of Hippo signaling caused by WWC3. Immunoprecipition was to examine WWC3 and LATS1 interaction. WWC3 knockdown drives a pronounced shift from the epithelial to the mesenchymal phenotype in lung cancer cells. In addition, WWC3 ectopic expression in lung cancer cells attenuates mesenchymal markers and increases the epithelial markers expressions; however, WWC3-ΔWW plasmid abrogated these effects. WWC3 silencing by shRNA exerts the opposite effect. Furthermore, WWC3 levels were inversely correlated with the levels of EMT inducers (Snail and Slug) in lung cancer cells and specimens. Immunoblotting revealed that WWC3 wild-type upregulates large tumor suppressor (LATS1) and yes-associated protein (YAP) phosphorylation through its WW domain, hence activating Hippo pathway. Knockdown of YAP and LATS1, as well as the as the Verteporfin (VP) usage, could reverse this effect caused by WWC3 silencing. These findings suggest that WWC3 works as a tumor suppressor to inhibit EMT process and confer its candidacy as a potential therapeutic target in lung cancer.