Fig. 2

Higher matrix stiffness alters malignant phenotypes and induces EMT occurrence in HCC cells independently. a Alteration in appearance and spreading area of HCC cells under higher stiffness stimulation. L, low-stiffness substrate, 6 KPa; M, medium-stiffness substrate, 10 KPa; H, high-stiffness substrate, 16 KPa. Scale bar, 1000 μm. b The mRNA expressions of MMP2, MMP9, SPP1, and CD44 in HCC cells grown on different stiffness substrates detected by qRT-PCR. c Analysis of cell movement under different stiffness stimulations by the real-time cell monitoring system. d Alexa-488 Phalloidin staining showed that higher matrix stiffness promotes F-actin polymerization in HCC cells (scale bar, 350 μm). e The expressions of N-cadherin, vimentin, α-SMA, and E-cadherin in HCC cells grown on different stiffness substrates in presence or absence of TGF β1. f Phosphorylation levels of Smad2 and Smad3 at different time points of TGFβ1 stimulation. g Snail expression in HCC cells grown on different stiffness substrates. h Expressions of E-cadherin, vimentin, and Snail in orthotopic HCC tumors in groups N, M, and H. N, normal stiffness; M, medium stiffness; H, high stiffness. Scale bar, 1000 μm. i Suppression of integrin β1 or α5 reversed the expressions of EMT markers in HCC cells grown on higher stiffness substrate. LV-shRNA-ITGβ1, lentivirus-shRNA-integrinβ1; LV-shRNA-ITGα5, lentivirus-shRNA-integrinα5. *P < 0.05, **P < 0.01, ***P < 0.001