Fig. 2

sFRP1 mediates the pro-tumoral effects of conditioned medium (CM) from NE-stimulated HSCs on malignant characteristics of HCC cells. a Compared with CM from NE-untreated LX2 cells, CM from NE-treated LX2 cells significantly enhanced the migration and invasion of HCC cells. b A decrease of E-cadherin expression and an increase of N-cadherin, vimentin, snail, c-Myc, cyclin D1, cancer stem cell markers of Nanog, and β-catenin were observed in HCC cells exposed to CM from NE-treated LX2. c NE (10 μM) significantly promoted the mRNA and protein expression of sFRP1 in HSCs (LX2 and p-HSC) compared with HCC cells, as analyzed by qRT-PCR. d Pretreated with prazosin (10 μM), 5-methylurapidi (5-Mu) (5 μM), propranolol (10 μM) or AMPK inhibitor dorsomorphin (0, 0.2, 1.0, 5.0 μM), LX-2 cells were treated with 10 μM NE. The expression of sFRP1 protein was detected by western blot analysis. e Compared with CM from NE-treated LX-2^{shRNA NC}, CM from NE-treated LX-2^{shRNA sFRP1} showed a significant decrease of invasion and migration of HCC cells in vitro, as measured by wound-healing migration assay and Matrigel invasion assay. f The expression levels of EMT markers (E-cadherin, N-cadherin, vimentin, and snail), proliferation-related gene cyclin D1, cancer stem cell marker Nanog, and β-catenin were measured by western blot analyses in HCC cells exposed to CM from NE-treated LX-2^{shRNA sFRP1} versus NE-treated LX-2^{shRNA NC} cells. g Immunofluorescent staining showed that β-catenin nuclear translocation was significantly decreased when HCC cells were exposed to CM from NE-treated LX-2^{shRNA sFRP1} versus NE-treated LX-2^{shRNA NC}. h Exogenous sFRP1 promoted the migration and invasion of HCC cells in vitro, as measured by wound-healing migration assay and Matrigel invasion assay. i A decrease in E-cadherin expression and an increase of N-cadherin, vimentin and snail expression, c-Myc, cyclin D1, Nanog and β-catenin were observed in HCC cells treated with exogenous sFRP1. *P < 0.05, **P < 0.01, *** P < 0.001