Department of Physiology, School of Medicine, Universidad Austral de Chile, Valdivia, Chile.
Center for Interdisciplinary Studies of the Nervous System (CISNe), Universidad Austral de Chile, Valdivia, Chile.
PLoS One. 2019 Feb 19;14(2):e0212321. doi: 10.1371/journal.pone.0212321. eCollection 2019.
Golgi phosphoprotein 3 (GOLPH3) is a conserved protein of the Golgi apparatus that in humans has been implicated in tumorigenesis. However, the precise function of GOLPH3 in malignant transformation is still unknown. Nevertheless, clinicopathological data shows that in more than a dozen kinds of cancer, including gliomas, GOLPH3 could be found overexpressed, which correlates with poor prognosis. Experimental data shows that overexpression of GOLPH3 leads to transformation of primary cells and to tumor growth enhancement. Conversely, the knocking down of GOLPH3 in GOLPH3-overexpressing tumor cells reduces tumorigenic features, such as cell proliferation and cell migration and invasion. The cumulative evidence indicate that GOLPH3 is an oncoprotein that promotes tumorigenicity by a mechanism that impact at different levels in different types of cells, including the sorting of Golgi glycosyltransferases, signaling pathways, and the actin cytoskeleton. How GOLPH3 connects mechanistically these processes has not been determined yet. Further studies are important to have a more complete understanding of the role of GOLPH3 as oncoprotein. Given the genetic diversity in cancer, a still outstanding aspect is how in this inherent heterogeneity GOLPH3 could possibly exert its oncogenic function. We have aimed to evaluate the contribution of GOLPH3 overexpression in the malignant phenotype of different types of tumor cells. Here, we analyzed the effect on cell migration that resulted from stable, RNAi-mediated knocking down of GOLPH3 in T98G cells of glioblastoma multiforme, a human glioma cell line with unique features. We found that the reduction of GOLPH3 levels produced dramatic changes in cell morphology, involving rearrangements of the actin cytoskeleton and reduction in the number and dynamics of focal adhesions. These effects correlated with decreased cell migration and invasion due to affected persistence and directionality of cell motility. Moreover, the knocking down of GOLPH3 also caused a reduction in autoactivation of focal adhesion kinase (FAK), a cytoplasmic tyrosine kinase that regulates focal adhesions. Our data support a model in which GOLPH3 in T98G cells promotes cell migration by stimulating the activity of FAK.
高尔基磷蛋白 3(GOLPH3)是高尔基体的一种保守蛋白,在人类中与肿瘤发生有关。然而,GOLPH3 在恶性转化中的精确功能仍然未知。尽管如此,临床病理数据表明,在包括神经胶质瘤在内的十多种癌症中,GOLPH3 都存在过度表达的情况,且与预后不良相关。实验数据表明,GOLPH3 的过表达会导致原代细胞的转化和肿瘤生长的增强。相反,在 GOLPH3 过表达的肿瘤细胞中敲低 GOLPH3,会降低肿瘤发生的特征,如细胞增殖、细胞迁移和侵袭。累积的证据表明,GOLPH3 是一种癌蛋白,通过影响不同类型细胞中不同水平的机制促进肿瘤发生,包括高尔基体糖基转移酶的分拣、信号通路和肌动蛋白细胞骨架。GOLPH3 如何在机制上连接这些过程尚未确定。进一步的研究对于更全面地了解 GOLPH3 作为癌蛋白的作用很重要。鉴于癌症的遗传多样性,一个仍然悬而未决的方面是,在这种固有的异质性中,GOLPH3 如何发挥其致癌功能。我们旨在评估 GOLPH3 过表达在不同类型肿瘤细胞恶性表型中的贡献。在这里,我们分析了在胶质母细胞瘤多形性(GBM)的 T98G 细胞中,通过稳定的 RNAi 介导的 GOLPH3 敲低对细胞迁移的影响,T98G 细胞是一种具有独特特征的人类神经胶质瘤细胞系。我们发现,GOLPH3 水平的降低会导致细胞形态发生剧烈变化,涉及肌动蛋白细胞骨架的重排和焦点黏附的数量和动态的减少。这些效应与细胞迁移和侵袭的减少有关,这是由于细胞运动的持久性和方向性受到影响。此外,GOLPH3 的敲低也导致细胞质酪氨酸激酶焦点黏附激酶(FAK)的自动激活减少,FAK 调节焦点黏附。我们的数据支持这样一种模型,即在 T98G 细胞中,GOLPH3 通过刺激 FAK 的活性促进细胞迁移。