Fig. 2

CRNDE is positively correlated with EGFR expression, thereby increasing proliferation and sorafenib resistance of HCC. A qRT-PCR analysis of CRNDE in various malignant compared with benign HCC cells (HepG2). B, C Western blot analysis of p-EGFR, EGFR, p-STAT3, and STAT3 protein levels in Hep3B cells (control and CRNDE depletion groups) and Huh7 cells (pcDNA3 and CRNDE overexpression groups). D, E qRT-PCR analysis of EGFR mRNA in Hep3B (control and CRNDE depletion groups) and Huh7 cells (pcDNA3 and CRNDE overexpression groups). F qRT-PCR analysis of CRNDE expression in 240 pairs of human HCC tissues (T) and adjacent normal (N) tissues. G qRT-PCR analysis of EGFR expression in 240 pairs of human HCC tissues (T) and adjacent normal (N) tissues. H Spearman correlation analysis of the clinical correlations between CRNDE and EGFR in HCC. I Representative IHC images of tumor xenografts stained with anti-Ki67 and anti-EGFR antibodies. Scale bar = 100 μm. J Huh7 cells were transfected with pcDNA3-control or CRNDE plasmids with siRNA-induced silencing of EGFR. Cell numbers were assessed after 48 h via the proliferation assay relative to control cells. K Huh7 cells transfected with pcDNA3-control and CRNDE overexpression plasmids with siRNA-induced silencing of EGFR were incubated with or without sorafenib and viability relative to control cells was assessed by MTT assay. L Western blot analysis of EGFR protein levels in Huh7 cells from pcDNA3-control and CRNDE overexpression groups with siRNA-induced silencing of EGFR. Statistical significance (p value) was calculated with the two-tailed Student's t-test for a single comparison between two groups. Data are presented as mean ± SD (*p < 0.05; **p < 0.01; ***p < 0.001)