[1]王红兵,张持,杨力,等.基底硬度对肝细胞胞间粘附复合物稳定性的影响[J].陆军军医大学学报(原第三军医大学学报),2013,35(20):2181-2186.
 Wang Hongbing,Zhang Chi,Yang Li,et al.Effects of substrate stiffness on stability of adhesion complexes of hepatic cells[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(20):2181-2186.
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基底硬度对肝细胞胞间粘附复合物稳定性的影响(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第20期
页码:
2181-2186
栏目:
论著
出版日期:
2013-10-30

文章信息/Info

Title:
Effects of substrate stiffness on stability of adhesion complexes of hepatic cells
作者:
王红兵张持杨力 邹小兵于光磊刘美霞许蓉林雨
重庆大学:生物工程学院,化学工程学院,数理工程学院
Author(s):
Wang Hongbing Zhang Chi Yang Li Zou Xiaobing Yu Guanglei Liu Meixia Xu Rong Lin Yu
College of Bioengineering, College of Chemistry and Chemical Engineering, College of Mathematical Engineering, Chongqing University, Chongqing, 400044, China
关键词:
肝细胞基底硬度粘附复合物β-连环蛋白
Keywords:
hepatic cells substrate stiffness adhesion complexes β-catenin
分类号:
R318;R322.41;R341
文献标志码:
A
摘要:
目的      通过比较汇合生长肝细胞(L02)对不同硬度基底(3.6、10 kPa和30 kPa)的响应,探查细胞间粘附复合物稳定性差异的成因。      方法      运用实时显微拍摄观察L02细胞在不同硬度基底上的形态特征;采用免疫荧光染色技术检测细胞肌动蛋白骨架构象和β-连环蛋白表达分布;应用Western blot方法检测β-连环蛋白的表达及其磷酸化水平。      结果      ①不同硬度基底上汇合生长L02细胞间粘附复合物稳定性不同,在乙二胺四乙酸(ethylene diamine tetraacetic acid, EDTA)作用下随基底硬度增加,细胞分离时间缩短;②汇合生长L02细胞在软基质上肌动蛋白骨架成皮质下环,β-连环蛋白分布于细胞连接处。随着基底硬度增大,细胞骨架环变得不完整,胞内骨架呈放射状分布,β-连环蛋白弥散分布于细胞质中;③随基底硬度增加L02细胞膜上β-连环蛋白表达显著降低(P<0.01),细胞质中磷酸化的p-β-连环蛋白(Tyr654)的表达量显著增加(P<0.01)。      结论      基底硬度影响肝细胞粘附复合物稳定性,基底硬度越大复合物稳定性越差,这与基底硬度增加导致的p-β-连环蛋白(Tyr654)表达量增高有关。
Abstract:
Objective      To investigate the effects of substrate stiffness on the stability of adhesion complexes of confluent hepatic (L02) cells cultured on substrates with different stiffness (3.6, 10, and 30 kPa) .       Methods      Real-time photomicrography was applied to record the morphology of L02 cells. Immunofluorescence staining was used to test the distribution of F-actin and β-catenin. Western blotting was applied to detect the expression of β-catenin and its phosphorylation.       Results      Adhesion complexes of confluent L02 cells exhibited different stability when these cells were cultured on substrates with different stiffness. When treated with EDTA, L02 cells on rigid substrate separated more quickly than those on soft substrate. F-actin of confluent L02 cells formed a ring-like conformation under the cortical, and β-catenin located at the cell-cell contact sites on soft substrate. With the increase of substrate stiffness, the ring-like cytoskeleton reduced and more F-actin distributed in cytoplasm, and β-catenin exhibited a similar distribution. As the substrate stiffness increased, the expression of β-catenin on membrane decreased (P<0.01) while that of phosphorylated β-catenin (p-β-catenin, Tyr654) in cytoplasm increased significantly (P<0.01).       Conclusion      Substrate stiffness affects the stability of adhesion complexes of confluent L02 cells observably. The cells on rigid substrate show weaker adhesion, which may be induced by up-regulation of p-β-catenin (Tyr654) expression.

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更新日期/Last Update: 2013-10-22