Matrix protease production, epithelial-to-mesenchymal transition marker expression and invasion of glioblastoma cells in response to osmotic or hydrostatic pressure
Matrix protease production, epithelial-to-mesenchymal transition marker expression and invasion of glioblastoma cells in response to osmotic or hydrostatic pressure
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DOI：
10.1038/s41598-020-59462-w
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发表时间：
2020-02-14
影响因子：
4.6
通讯作者：
Parat, Marie-Odile
中科院分区：
文献类型：
--
作者：
Pu, Wenjun;Qiu, Jiawen;Parat, Marie-Odile
肿瘤微环境中的静水压和渗透压均发生改变。胶质母细胞瘤（GBM）是一种侵袭性高、预后差的脑肿瘤。我们假设物理力和渗透力调节胶质母细胞瘤（GBM）的侵袭性。使用氯化钠或水调节GBM细胞培养基的渗透压。或者，细胞受到增加的静水力。使用酶谱法和实时 qPCR 研究蛋白水解谱和上皮间质转化 (EMT)。评估的 EMT 标志物包括 Snail-1、Snail-2、N-钙粘蛋白、Twist 和波形蛋白。使用细胞外基质包被的 Transwell 插入物在体外研究侵袭。响应渗透压和机械压力，GBM 细胞系 U87 和 U251 以及患者来源的神经癌球上调了尿激酶型纤溶酶原激活剂 (uPA) 和/或基质金属蛋白酶 (MMP) 以及一些测试的 EMT 标志物的表达。当置于渗透压增加的培养基中时，贴壁细胞系侵入更多。因此，GBM 通过增加基质降解酶的产生并采用类似于 EMT 的表型来响应渗透压或机械压力。更好地了解压力增加促进 GBM 侵袭性的分子和细胞机制可能有助于开发创新的治疗方法。
Both hydrostatic and osmotic pressures are altered in the tumour microenvironment. Glioblastoma (GBM) is a brain tumour with high invasiveness and poor prognosis. We hypothesized that physical and osmotic forces regulate glioblastoma (GBM) invasiveness. The osmotic pressure of GBM cell culture medium was adjusted using sodium chloride or water. Alternatively, cells were subjected to increased hydrostatic force. The proteolytic profile and epithelial-mesenchymal transition (EMT) were investigated using zymography and real-time qPCR. The EMT markers assessed were Snail-1, Snail-2, N-cadherin, Twist and vimentin. Invasion was investigated in vitro using extracellular matrixcoated Transwell inserts. In response to osmotic and mechanical pressure, GBM cell lines U87 and U251 and patient-derived neural oncospheres upregulated the expression of urokinase-type plasminogen activator (uPA) and/or matrix metalloproteinases (MMPs) as well as some of the EMT markers tested. The adherent cell lines invaded more when placed in media of increased osmolality. Therefore, GBM respond to osmotic or mechanical pressure by increasing matrix degrading enzyme production, and adopting a phenotype reminiscent of EMT. Better understanding the molecular and cellular mechanisms by which increased pressure promotes GBM invasiveness may help to develop innovative therapeutic approaches.