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

SOX2 silencing represses tumor growth in mice with HCC by activating CCAT1, EGFR, miR-222-5p and by downregulating CYLD. HepG2 cells expressing sh-SOX2, miR-222-5p agomir or corresponding controls (sh-NC or agomir-NC) were administrated subcutaneously to nude mice for developing in vivo tumor model. (A) Tumor growth rate in nude mice. (B) Representative images showing tumor mass collected from nude mice at day 30. (C) Tumor weights in nude mice. (D) mRNA expression of SOX2, EGFR and CYLD and CCAT1 and miR-222-5p expression in nude mice xenografts determined by RT-qPCR normalized to U6 and β-actin. (E, F) Protein expression of SOX2, EGFR and CYLD in nude mice xenografts determined by western blot analysis normalized to GAPDH. n = 6/group. * p vs. untreated mice; # p vs. mice treated with HepG2 cells expressing sh-SOX2 + agomir NC. Data were 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 testing.

Figure 7. SOX2 silencing represses tumor growth in mice with HCC by activating CCAT1, EGFR, miR-222-5p and by downregulating CYLD. HepG2 cells expressing sh-SOX2, miR-222-5p agomir or corresponding controls (sh-NC or agomir-NC) were administrated subcutaneously to nude mice for developing in vivo tumor model. (A) Tumor growth rate in nude mice. (B) Representative images showing tumor mass collected from nude mice at day 30. (C) Tumor weights in nude mice. (D) mRNA expression of SOX2, EGFR and CYLD and CCAT1 and miR-222-5p expression in nude mice xenografts determined by RT-qPCR normalized to U6 and β-actin. (E, F) Protein expression of SOX2, EGFR and CYLD in nude mice xenografts determined by western blot analysis normalized to GAPDH. n = 6/group. * p < 0.05 vs. untreated mice; # p < 0.05 vs. mice treated with HepG2 cells expressing sh-SOX2 + agomir NC. Data were 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 testing.