Náger Mireia, Santacana Maria, Bhardwaj Deepshikha, Valls Joan, Ferrer Isidre, Nogués Pere, Cantí Carles, Herreros Judit
a Departments of Basic Medical Sciences & Experimental Medicine ; University of Lleida & IRBLleida ; Lleida , Spain.
b Immunohistochemical and Biostatistics and Epidemiology Units; IRBLleida ; Lleida , Spain.
Cell Cycle. 2015;14(22):3644-55. doi: 10.1080/15384101.2015.1104443.
Glioblastoma multiforme (GBM) is a fast growing brain tumor characterized by extensive infiltration into the surrounding tissue and one of the most aggressive cancers. GBM is the most common glioma (originating from glial-derived cells) that either evolves from a low grade astrocytoma or appears de novo. Wnt/β-catenin and Hepatocyte Growth Factor (HGF)/c-Met signaling are hyperactive in human gliomas, where they regulate cell proliferation, migration and stem cell behavior. We previously demonstrated that β-catenin is phosphorylated at Y142 by recombinant c-Met kinase and downstream of HGF signaling in neurons. Here we studied phosphoY142 (PY142) β-catenin and dephospho S/T β-catenin (a classical Wnt transducer) in glioma biopsies, GBM cell lines and biopsy-derived glioma cell cultures. We found that PY142 β-catenin mainly localizes in the nucleus and signals through transcriptional activation in GBM cells. Tissue microarray analysis confirmed strong nuclear PY142 β-catenin immunostaining in astrocytoma and GBM biopsies. By contrast, active β-catenin showed nuclear localization only in GBM samples. Western blot analysis of tumor biopsies further indicated that PY142 and active β-catenin accumulate independently, correlating with the expression of Snail/Slug (an epithelial-mesenchymal transition marker) and Cyclin-D1 (a regulator of cell cycle progression), respectively, in high grade astrocytomas and GBMs. Moreover, GBM cells stimulated with HGF showed increasing levels of PY142 β-catenin and Snail/Slug. Importantly, the expression of mutant Y142F β-catenin decreased cell detachment and invasion induced by HGF in GBM cell lines and biopsy-derived cell cultures. Our results identify PY142 β-catenin as a nuclear β-catenin signaling form that downregulates adhesion and promotes GBM cell invasion.
多形性胶质母细胞瘤(GBM)是一种生长迅速的脑肿瘤,其特征是广泛浸润周围组织,是最具侵袭性的癌症之一。GBM是最常见的胶质瘤(起源于神经胶质衍生细胞),它要么由低级别星形细胞瘤演变而来,要么是新发的。Wnt/β-连环蛋白和肝细胞生长因子(HGF)/c-Met信号通路在人类胶质瘤中过度活跃,它们在其中调节细胞增殖、迁移和干细胞行为。我们之前证明,β-连环蛋白在神经元中被重组c-Met激酶在Y142位点磷酸化,且处于HGF信号通路的下游。在此,我们研究了胶质瘤活检组织、GBM细胞系以及活检来源的胶质瘤细胞培养物中的磷酸化Y142(PY142)β-连环蛋白和去磷酸化的丝氨酸/苏氨酸β-连环蛋白(一种经典的Wnt转导蛋白)。我们发现,PY142 β-连环蛋白主要定位于细胞核,并通过GBM细胞中的转录激活发出信号。组织芯片分析证实,星形细胞瘤和GBM活检组织中存在强烈的细胞核PY142 β-连环蛋白免疫染色。相比之下,活性β-连环蛋白仅在GBM样本中显示细胞核定位。肿瘤活检组织的蛋白质印迹分析进一步表明,PY142和活性β-连环蛋白独立积累,分别与高级别星形细胞瘤和GBM中Snail/Slug(一种上皮-间质转化标志物)和细胞周期蛋白D1(细胞周期进程的调节因子)的表达相关。此外,用HGF刺激的GBM细胞显示PY142 β-连环蛋白和Snail/Slug水平升高。重要的是,突变型Y142F β-连环蛋白的表达降低了GBM细胞系和活检来源的细胞培养物中HGF诱导的细胞脱离和侵袭。我们的研究结果确定PY142 β-连环蛋白是一种细胞核β-连环蛋白信号形式,它下调黏附并促进GBM细胞侵袭。