Effects of substrate stiffness on confluent growth of hepatic and hepatoma carcinoma cells
- VernacularTitle:基底硬度对肝细胞和肝癌细胞融合生长的影响
- Author:
Rong ZHANG
1
;
Hong-bing WANG
1
;
Yan-zi YANGBEN
1
;
Qiao-yan TAN
1
;
Li YANG
1
Author Information
1. Collge of Bioengieering, Chongqing University
- Publication Type:Journal Article
- Keywords:
Hepatic cells;
Hepatoma carcinoma cells;
Substrate stiffness;
E-cadherin distribution;
Src kinase activity
- From:
Journal of Medical Biomechanics
2013;28(1):E091-E096
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the cause of differences in confluent growth between hepatic(L02) and hepatoma carcinoma(HCCLM3) cells by comparing responses of the two cells to different substrate stiffness (0.5, 4 kPa and glass). MethodsThe real-time photomicrography, immunofluorescence staining, flow cytometry, and Western Blotting techniques were respectively employed to observe the morphological characteristics, the cytoskeleton conformation and the distribution of E-cad of confluent L02 and HCCLM3 cells on different substrates, and test the changes in expression of E-cad, Integrinβ1 and p-Src. Results (1) Confluent L02 cells displayed a round or cubic shape, while HCCLM3 cells showed a polygon shape. The morphology of HCCLM3 cells were spread and polarized more obviously than that of L02 cells. With the increase of substrate stiffness, the variation of L02 cells with time was smaller than that of HCCLM3 cells. (2) The cytoskeleton of confluent L02 cells showed a ring-like conformation under the cortex, and E-cad was located at the cell-cell contact sites. However, the ring-like cytoskeleton of HCCLM3 cells was incomplete and distributed radially along the basement, while E-cad was dispersed in cytoplasm. (3) As the substrate stiffness increased, expression of E-cadherin in both L02 and HCCLM3 cells was significantly decreased (P<0.01), while the level of p-Src and integrinβ1 was increased significantly, with greater changes in HCCLM3 cells than in L02 cells. Conclusions The assembling of cortical ring-like cytoskeleton was positively correlated with the location of E-cad at the cell-cell contact sites. The substrate stiffness showed a more obvious impact on the balanced regulation between cadherin and integrin mediated adhesion system of hepatocarcinoma cells than that of hepatic cells.
