Transfection of C/EBPα-saRNA in SMMC-7721 and HepG2 cells resulted in inhibition of migration.

<p>(A–B) Gene expression of C/EBPα (A) and albumin (B) in SMMC-7721 and HepG2 cells transfected with C/EBPα-saRNA or scramble-saRNA as the control for 48 h. (C) Transwell assays show reduced migration of SMMC-7721 and HepG2 cells transfected with C/EBPα-saRNA compared with the control after 18 h. Magnification, 100×. Data represent mean±SD.</p

Primers and product size.

<p>Primers and product size.</p

Intravenous injection of C/EBPα-saRNA-dendrimer inhibited tumor metastasis and improved liver function in nude mice with tumors from HepG2-RFP cells.

<p>(A) Imaging of liver orthotopic xenograft tumors after injection of C/EBPα-saRNA on day 40 post-injection of HepG2-RFP cells and scramble-saRNA as the control. Higher fluorescence intensity and intrahepatic metastasis was observed in the control group. (B) C/EBPα-saRNA-dendrimer was tested for nuclease sensitivity in nude mice for indicated times. (C) Survival analysis of C/EBPα-saRNA-injected mouse groups and the control (n = 9) (log rate: 0.0167). (D) Statistical table of tumor formation, intrahepatic and distant metastasis. (E) Hematoxylin and eosin staining of liver and lung from C/EBPα-saRNA injected mice and the control. (F) Expression of C/EBPα was determined by RT-PCR and western blotting in liver tissue of mice injected with C/EBPα-saRNA-dendrimer or the control. (G-I) C/EBPα-saRNA-dendrimer injected nude mice showed significant changes in circulating levels of serum albumin (G), AST (H) and ALT (I). Data represent mean±SD.</p

Transfection of C/EBPα-saRNA inhibits metastasis by targeting EGFR/β-catenin signaling pathway.

<p>(A) C/EBPα binding sites in EGFR and β-catenin genomic regions. (B) Western blot shows decreased ADAM17, EGFR protein and phosphorylation levels and decreased β-catenin in hepatoma cells transfected with C/EBPα-saRNA. (C) Reduced gene expression of c-Myc, Axin2, CCDN1, and Lgr5 in C/EBPα-saRNA transfected HepG2 cells compared with scramble-saRNA transfected as the control. (D) Immunohistochemistry analysis of EGFR, phosphorylation of EGFR, and β-catenin in liver tissues. (E) Western blot shows inhibited ADAM17, EGFR protein and phosphorylation levels and suppressed β-catenin in liver tissue of C/EBPα-saRNA-dendrimer-treated mice (n = 9).</p

Transfection of C/EBPα-saRNA in hepatoma cells inhibited migration, invasion and EMT.

<p>(A-B) Wound healing assays of SMMC-7721 (A) and HepG2 (B) cells transfected with scramble-saRNA or C/EBPα-saRNA at 0 h, 12 h, and 24 h. Magnification, 40×. (C) Transwell assays show suppressed invasion of cells transfected with C/EBPα-saRNA compared with scramble-saRNA as the control after 24 h. Magnification, 100×. (D-E) Western blot analysis shows decreased N-cadherin, Slug, and Vimentin and up-regulation of E-cadherin, albumin, and C/EBPα in hepatoma cells transfected with C/EBPα-saRNA compared with the control. Data represent mean±SD.</p