Relative Rigidity of Cell–Substrate Effects on Hepatic and Hepatocellular Carcinoma Cell Migration

Abstract

Polyacrylamide gels with different stiffness and glass were employed as substrates to investigate how substrate stiffness affects the cellular stiffness of adherent hepatocellular carcinoma (HCCLM3) and hepatic (L02) cells. The interaction of how cell-substrate stiffness influences cell migration was also explored. An atom force microscope measured the stiffness of HCCLM3 and L02 cells on different substrates. Further, F-actin assembly was analyzed using immunofluorescence and Western blot. Finally, cell-surface expression of integrin β1 was quantified by flow cytometry. The results show that, while both HCCLM3 and L02 cells adjusted their cell stiffness to comply with the stiffness of the substrate they were adhered to, their tuning capabilities were different. HCCLM3 cell stiffness complied when substrate stiffness was between 1.1 and 33.7 kPa, whereas the analogous stiffness for L02 cells occurred at a higher substrate stiffness, 3.6 kPa up to glass. These ranges correlated with F-actin filament assembly and integrin β1 expression. In a migration assay, HCCLM3 cells migrated faster on a relatively soft substrate, while L02 cells migrated faster on substrates that were relatively rigid. These findings indicate that different tuning capabilities of HCCLM3 and L02 cells may influence cell migration velocity on substrates with different stiffness by regulating cy- toskeleton remodeling and integrin β1 expression.

Keywords:

• Relative cell–substrate stiffness
• cytoskeleton assembly
• integrin β1 expression
• cell migration
• adaptation range
• hepatic carcinoma cells

Acknowledgements

This work was supported in part by grants from the National Nature Science Foundation of China (Nos 11032012, 30870608 and 10472137), the Innovation and Attracting Talents Program for College and University (‘111’ Project) (B06023), the Key Science and Technology Program of CQ CSTC (CSTC, 2009AA5045) and the sharing fund of Chongqing University’s large-scale equipment.