Lee Yeonju, Koay Eugene J, Zhang Weijia, Qin Lidong, Kirui Dickson K, Hussain Fazle, Shen Haifa, Ferrari Mauro
Department of Nanomedicine, Houston Methodist Research Institute, Houston, Texas, United States of America.
Department of Nanomedicine, Houston Methodist Research Institute, Houston, Texas, United States of America; Department of Radiation Oncology, M. D. Anderson Cancer Center, Houston, Texas, United States of America.
PLoS One. 2014 Oct 14;9(10):e107973. doi: 10.1371/journal.pone.0107973. eCollection 2014.
We report cell mechanical changes in response to alteration of expression of the human equilibrative nucleoside transporter-1 (hENT1), a most abundant and widely distributed plasma membrane nucleoside transporter in human cells and/or tissues. Modulation of hENT1 expression level altered the stiffness of pancreatic cancer Capan-1 and Panc 03.27 cells, which was analyzed by atomic force microscopy (AFM) and correlated to microfluidic platform. The hENT1 knockdown induced reduction of cellular stiffness in both of cells up to 70%. In addition, cellular phenotypic changes such as cell morphology, migration, and expression level of epithelial-mesenchymal transition (EMT) markers were observed after hENT1 knockdown. Cells with suppressed hENT1 became elongated, migrated faster, and had reduced E-cadherin and elevated N-cadherin compared to parental cells which are consistent with epithelial-mesenchymal transition (EMT). Those cellular phenotypic changes closely correlated with changes in cellular stiffness. This study suggests that hENT1 expression level affects cellular phenotype and cell elastic behavior can be a physical biomarker for quantify hENT1 expression and detect phenotypic shift. Furthermore, cell mechanics can be a critical tool in detecting disease progression and response to therapy.
我们报告了人类平衡核苷转运体-1（hENT1）表达改变时细胞的力学变化，hENT1是人类细胞和/或组织中最丰富且分布广泛的质膜核苷转运体。通过原子力显微镜（AFM）分析并与微流控平台相关联，hENT1表达水平的调节改变了胰腺癌Capan-1和Panc 03.27细胞的硬度。hENT1基因敲低导致两种细胞的细胞硬度降低高达70%。此外，在hENT1基因敲低后，观察到细胞表型变化，如细胞形态、迁移以及上皮-间质转化（EMT）标志物的表达水平。与亲代细胞相比，hENT1受抑制的细胞变得细长，迁移更快，E-钙黏蛋白减少，N-钙黏蛋白升高，这与上皮-间质转化（EMT）一致。这些细胞表型变化与细胞硬度的变化密切相关。本研究表明，hENT1表达水平影响细胞表型，细胞弹性行为可作为量化hENT1表达和检测表型转变的物理生物标志物。此外，细胞力学可成为检测疾病进展和治疗反应的关键工具。
