CXCR4 is involved in proliferation, EMT, and stemness characteristics induced by matrix stiffness. mRNA (A) and protein (B) were significantly up-regulated with increasing matrix stiffness. (C) Expression levels of CXCR4 in HCC tissues with normal (groups L, N=25), medium (group M, N=35), and high liver stiffness backgrounds (group H, N=46). (D) Hep3B and Huh7 cells transfected with shRNA were subjected to Western blotting for CXCR4 expression. (E) MTT revealed that stiffness-mediated HCC proliferation was abrogated by CXCR4 gene deletion. HCC cells seeded on low or high stiffness were transduced with NT shRNA or CXCR4 shRNA. Stiffness-mediated HCC (PCNA) and CyclinD1 expression (F), EMT (G) and stemness characteristics (H) were abrogated by CXCR4 knockdown. Bar: 70μM. *P < 0.05, **P < 0.01.