School of Life Science and Technology, Harbin Institute of Technology, Harbin 150001, China.
College of Life Science, China West Normal University, Nanchong 637002, China.
Int J Mol Sci. 2020 Feb 17;21(4):1349. doi: 10.3390/ijms21041349.
In order to study the effect of stress changes on cell adhesion, HUVEC, and MCF-7 cells were treated with simulated microgravity effect (SMG) and overloading (OL).
Rotating Wall Vessel (2D-RWVS) bioreactor was used to create different culture conditions. In addition, the alteration of cell adhesion states, adhesion proteins, and relating factors of adhesion molecules under these two conditions were detected using cell adhesion assay, immunofluorescence, western blot, and qRT-PCR technology.
The results showed that the adhesion of cells decreased under SMG, while increased under OL. The expressions of integrin β1, paxillin, and E-cadherin under SMG condition were down-regulated as compared to that of the control group showing a time-dependent pattern of the decreasing. However, under OL condition, the expressions of adhesion proteins were up-regulated as compared to that of the control group, with a time-dependent pattern of increasing. EMT transcription factors Snail, twist, and ZEB1 were up-regulated under SMG while down-regulated under OL.
Collectively our results indicated that cells could respond to stress changes to regulate the expressions of adhesion proteins and adapt their adhesion state to the altered mechanical environment. The altered cell adhesion in response to the mechanical stress may involve the changed expression of EMT-inducing factors, Snail, Twist, and ZEB1under the SMG/OL conditions.
为了研究细胞黏附力受应力变化的影响,本研究采用模拟微重力(SMG)和过载(OL)两种方法对 HUVEC 和 MCF-7 细胞进行处理。
采用旋转壁式生物反应器(2D-RWVS)创建不同的培养条件。采用细胞黏附实验、免疫荧光、Western blot 和 qRT-PCR 技术检测两种条件下细胞黏附状态、黏附蛋白以及黏附分子相关因子的变化。
结果显示,SMG 下细胞黏附减少,OL 下细胞黏附增加。与对照组相比,SMG 条件下整合素β1、桩蛋白和 E-钙黏蛋白的表达呈时间依赖性下调。然而,OL 条件下,黏附蛋白的表达呈时间依赖性上调。SMG 下 EMT 转录因子 Snail、Twist 和 ZEB1 上调,OL 下下调。
总之,我们的研究结果表明,细胞可以对压力变化做出反应,调节黏附蛋白的表达,并使其黏附状态适应改变的机械环境。在 SMG/OL 条件下,细胞对机械应激的改变黏附可能涉及 EMT 诱导因子(如 Snail、Twist 和 ZEB1)表达的改变。