Research Paper Volume 13, Issue 8 pp 12207—12223

Anti-oncogenic effects of SOX2 silencing on hepatocellular carcinoma achieved by upregulating miR-222-5p-dependent CYLD via the long noncoding RNA CCAT1

EGFR induces cell proliferation, migration and invasion via miR-222-5p upregulation in HepG2 cells. (A) Representative immunohistochemistry micrographs showing EGFR expression in normal liver (n = 28), para-cancerous (n = 68) and HCC tissues (n = 68) (400 ×). (B) EGFR positive expression in normal liver (n = 28), para-cancerous (n = 68) and HCC tissues (n = 68) determined by immunohistochemistry. (C) EGFR mRNA expressions in human HCC cell lines detected by RT-qPCR normalized to β-actin. HepG2 cells were treated with oe-NC, oe-EGFR-1, oe-EGFR-2, oe-EGFR-3, DMSO or EGFR-inhibitor. (D) EGFR expression in HepG2 cells detected by RT-qPCR. (E) miR-222-5p expression in HepG2 cells detected by RT-qPCR normalized to U6. (F) Cell viability determined by MTT assay. (G, H) Cell migration determined by Transwell assay (200 ×). (I, J) Cell invasion determined by Transwell assay (200 ×). * p vs. normal liver cell, LO2 or HepG2 cells treated with oe-NC; # p vs. HepG2 treated with DMSO. Data are expressed as mean ± standard deviation. Data from multiple groups were compared by one-way ANOVA and Tukey’s post hoc test. Data were compared between groups at different time points by repeated measures ANOVA and Bonferroni post hoc test.

Figure 3. EGFR induces cell proliferation, migration and invasion via miR-222-5p upregulation in HepG2 cells. (A) Representative immunohistochemistry micrographs showing EGFR expression in normal liver (n = 28), para-cancerous (n = 68) and HCC tissues (n = 68) (400 ×). (B) EGFR positive expression in normal liver (n = 28), para-cancerous (n = 68) and HCC tissues (n = 68) determined by immunohistochemistry. (C) EGFR mRNA expressions in human HCC cell lines detected by RT-qPCR normalized to β-actin. HepG2 cells were treated with oe-NC, oe-EGFR-1, oe-EGFR-2, oe-EGFR-3, DMSO or EGFR-inhibitor. (D) EGFR expression in HepG2 cells detected by RT-qPCR. (E) miR-222-5p expression in HepG2 cells detected by RT-qPCR normalized to U6. (F) Cell viability determined by MTT assay. (G, H) Cell migration determined by Transwell assay (200 ×). (I, J) Cell invasion determined by Transwell assay (200 ×). * p < 0.05 vs. normal liver cell, LO2 or HepG2 cells treated with oe-NC; # p < 0.05 vs. HepG2 treated with DMSO. Data are expressed as mean ± standard deviation. Data from multiple groups were compared by one-way ANOVA and Tukey’s post hoc test. Data were compared between groups at different time points by repeated measures ANOVA and Bonferroni post hoc test.