School of Biomedical Engineering, Southern Medical University, Guangzhou, China.
Laboratory of Biophysics, School of Medicine, South China University of Technology, Guangzhou, China.
Arch Biochem Biophys. 2022 Aug 15;725:109281. doi: 10.1016/j.abb.2022.109281. Epub 2022 May 7.
Cervical cancer microenvironment is involved in the regulation of the behavior, morphology, and mechanical properties of the cervical cancer cells. Integrins expressed on the cell membrane combine with the factors of the microenvironment to determine cervical cancer cells' properties. The mechanical properties of integrin-extracellular matrix (ECM) interactions are important for the mechanotransduction of cervical cancer cells. However, the quantified study on the adhesion force and binding probabilities between collagen and integrins on cervical cancer cells grown on different stiffness substrates have not been reported.
Polyacrylamide (PA) gel was used as substrate to mimic the mechanical microenvironment of cancer cells. ImageJ software was used to measure the perimeter and area of the cells. SiHa cells were stained with FITC phalloidine to observe the cytoskeleton. Atomic force microscopy (AFM) was used to measure the cell mechanical properties.
The perimeters of SiHa cells grown on different stiffness substrates were 63.4 ± 1.3, 102.8 ± 4.0, and 152.6 ± 4.1 μm, for soft, intermediate, and stiff substrates, respectively. These areas were 277.2 ± 13.3, 428.9 ± 26.0, and 1166.0 ± 63.2 μm, for soft, intermediate, and stiff substrates, respectively. The Young's modulus of SiHa cells grown on stiff substrates (3.0 ± 0.02 kPa) was higher compared with those on soft substrates (0.6 ± 0.01 kPa) or intermediate substrates (bimodal distribution, 1.36 and 1.67 kPa). The adhesion force between the functionalized probe and SiHa cells grown on glass (55.65 ± 0.78 pN) was significantly greater than that on stiff (47.03 ± 0.85 pN), intermediate (34.07 ± 0.58 pN) and soft (27.94 ± 0.47 pN) substrates. The binding probabilities of the collagen-integrin of the four rigid substrates were significantly differed.
The changes in substrate stiffness can obviously regulate SiHa cells' mechanical properties, such as the Young's modulus. The adhesion force and binding probabilities of SiHa cells both increased with increasing substrate strength.
宫颈癌微环境参与调节宫颈癌细胞的行为、形态和力学特性。细胞膜上表达的整合素与微环境中的因子结合,决定宫颈癌细胞的特性。整合素-细胞外基质(ECM)相互作用的力学特性对于宫颈癌细胞的力学转导非常重要。然而,关于在不同硬度基底上生长的宫颈癌细胞与胶原蛋白和整合素之间的粘附力和结合概率的定量研究尚未报道。
使用聚丙烯酰胺(PA)凝胶作为基底来模拟癌细胞的力学微环境。ImageJ 软件用于测量细胞的周长和面积。用 FITC 鬼笔环肽对 SiHa 细胞进行染色以观察细胞骨架。原子力显微镜(AFM)用于测量细胞力学特性。
在不同硬度基底上生长的 SiHa 细胞的周长分别为 63.4±1.3、102.8±4.0 和 152.6±4.1μm,对应的基底分别为软、中和硬。这些面积分别为 277.2±13.3、428.9±26.0 和 1166.0±63.2μm,对应的基底分别为软、中和硬。在硬基底上生长的 SiHa 细胞的杨氏模量(3.0±0.02kPa)高于在软基底(0.6±0.01kPa)或中硬基底(双峰分布,1.36 和 1.67kPa)上生长的细胞。功能化探针与在玻璃上生长的 SiHa 细胞(55.65±0.78pN)之间的粘附力明显大于在硬基底(47.03±0.85pN)、中硬基底(34.07±0.58pN)和软基底(27.94±0.47pN)上的粘附力。四种刚性基底上胶原蛋白-整合素的结合概率有显著差异。
基底硬度的变化可以明显调节 SiHa 细胞的力学特性,如杨氏模量。SiHa 细胞的粘附力和结合概率都随基底强度的增加而增加。