CD44v6+ Hepatocellular Carcinoma Cells Maintain Stemness Properties through Met/cJun/Nanog Signaling

Figure 2

The HGF/MET signaling pathway contributes to maintenance of stemness in CD44v6+ HCC Cells. (a) Western blot analysis of the Met protein levels in 18 paired HCC tissues and adjacent nontumor tissues selected randomly. β-Actin was used as a normalized control. (b) Kaplan–Meier survival analysis of overall survival were compared according to the expression levels of the Met in HCC tissues. Patients with Met expression median months vs. 48 months, log-rank test, , . The immunohistochemical staining shows the expressions of Met in surgical specimen from patients with HCC. The expression of Met in tumor tissue was significantly higher than in adjacent nontumor tissue. Scale bar, 20 μm. (c) The expression of EMT-related genes, including Snail1, Slug, and Twist in CD44v6+ and CD44v6- cells. β-Actin was used as a normalized control. (d) The expression of Met and the stemness relative genes, including Nanog, Sox2, and Oct4 in CD44v6+ and CD44v6- cells. β-Actin was used as a normalized control. (e) shMet CD44v6+ cells and scrambled cells were subcutaneously injected into the BALB/c mice ( in each group). Black arrow means NC group, red arrow means Met shRNA group. The tumor volume from each group was tested. For statistical analysis, , , -test. The representative images of IHC staining of Met in NC and shMet group tumor tissues. Bars: 200 μm. (f) The representative images of spheres and histogram analysis in indicated cells. Scale bar, 200 μm. , -test. (g) The representative images of the wound-healing experiment and histogram analysis in indicated cells. Scale bar, 100 μm. , -test. (h) Representative images of transwell migration and invasion in indicated cells. Scale bar, 200 μm. , , -test.

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