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 downregulation inhibits cell proliferation, migration, and invasion by downregulating miR-222-5p-targeted CYLD in HepG2 cells. HepG2 cells were treated with sh-NC + mimic-NC, sh-EGFR + mimic-NC, sh-EGFR + miR-222-5p mimic, sh-NC + si-NC, sh-EGFR + si-NC, sh-EGFR + si-CYLD, inhibitor-NC + si-NC, miR-222-5p inhibitor + si-NC, or miR-222-5p inhibitor + si-CYLD. (A) mRNA expression of miR-222-5p and CYLD in HepG2 cells detected by RT-qPCR normalized to U6 and β-actin. (B) Cell viability determined by MTT assay. (C, D) Cell migration determined by Transwell assay (200 ×). (E, F) Cell invasion determined by Transwell assay (200 ×). * p vs. HepG2 treated with sh-NC + mimic-NC, inhibitor-NC + si-NC, or sh-NC + si-NC. # p vs. HepG2 treated with sh-EGFR + mimic-NC, sh-EGFR + si-NC, or miR-222-5p inhibitor + si-NC. 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 5. EGFR downregulation inhibits cell proliferation, migration, and invasion by downregulating miR-222-5p-targeted CYLD in HepG2 cells. HepG2 cells were treated with sh-NC + mimic-NC, sh-EGFR + mimic-NC, sh-EGFR + miR-222-5p mimic, sh-NC + si-NC, sh-EGFR + si-NC, sh-EGFR + si-CYLD, inhibitor-NC + si-NC, miR-222-5p inhibitor + si-NC, or miR-222-5p inhibitor + si-CYLD. (A) mRNA expression of miR-222-5p and CYLD in HepG2 cells detected by RT-qPCR normalized to U6 and β-actin. (B) Cell viability determined by MTT assay. (C, D) Cell migration determined by Transwell assay (200 ×). (E, F) Cell invasion determined by Transwell assay (200 ×). * p < 0.05 vs. HepG2 treated with sh-NC + mimic-NC, inhibitor-NC + si-NC, or sh-NC + si-NC. # p < 0.05 vs. HepG2 treated with sh-EGFR + mimic-NC, sh-EGFR + si-NC, or miR-222-5p inhibitor + si-NC. 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.