BIOMATEN, Middle East Technical University (METU), Center of Excellence in Biomaterials and Tissue Engineering, Ankara, 06800, Turkey.
Department of Biotechnology, Middle East Technical University (METU), Ankara, 06800, Turkey.
Adv Biol (Weinh). 2021 Jan;5(1):e2000048. doi: 10.1002/adbi.202000048. Epub 2020 Dec 23.
Mechanotransduction proteins transfer mechanical stimuli through nucleo-cytoskeletal coupling and affect the nuclear morphology of cancer cells. However, the contribution of actin filament integrity has never been studied directly. It is hypothesized that differences in nuclear deformability of cancer cells are influenced by the integrity of actin filaments. In this study, transparent micropatterned surfaces as simple tools to screen cytoskeletal and nuclear distortions are presented. Surfaces decorated with micropillars are used to culture and image breast cancer cells and quantify their deformation using shape descriptors (circularity, area, perimeter). Using two drugs (cytochalasin D and jasplakinolide), actin filaments are disrupted. Deformation of cells on micropillars is decreased upon drug treatment as shown by increased circularity. However, the effect is much smaller on benign MCF10A than on malignant MCF7 and MDAMB231 cells. On micropatterned surfaces, molecular analysis shows that Lamin A/C and Nesprin-2 expressions decreased but, after drug treatment, increased in malignant cells but not in benign cells. These findings suggest that Lamin A/C, Nesprin-2 and actin filaments are critical in mechanotransduction of cancer cells. Consequently, transparent micropatterned surfaces can be used as image analysis platforms to provide robust, high throughput measurements of nuclear deformability of cancer cells, including the effect of cytoskeletal elements.
力学转导蛋白通过核-细胞骨架偶联将机械刺激传递,并影响癌细胞的核形态。然而, actin 丝完整性的作用从未被直接研究过。假设癌细胞核变形的差异受 actin 丝完整性的影响。在这项研究中,透明微图案化表面作为筛选细胞骨架和核变形的简单工具被提出。用微柱装饰的表面用于培养和成像乳腺癌细胞,并使用形状描述符(圆度、面积、周长)量化它们的变形。使用两种药物(细胞松弛素 D 和 jasplakinolide)破坏 actin 丝。药物处理后,细胞在微柱上的变形减少,表现为圆度增加。然而,这种效果在良性 MCF10A 细胞上比在恶性 MCF7 和 MDAMB231 细胞上要小得多。在微图案化表面上,分子分析表明 Lamin A/C 和 Nesprin-2 的表达减少,但在药物处理后,恶性细胞的表达增加,而良性细胞则没有。这些发现表明 Lamin A/C、Nesprin-2 和 actin 丝在癌细胞的力学转导中是至关重要的。因此,透明微图案化表面可用作图像分析平台,提供对癌细胞核变形的稳健、高通量测量,包括细胞骨架元件的影响。