Figure 1.
IHC staining of ZNF217 in CRC.
(A-Left) Cytoplasma staining of ZNF217 in CRC cells. (A-Right) Lack of ZNF217 expression in normal colonic epithelial cells. (B) The percentage of positively stained cells in cancer and adjacent normal tissues (*P < 0.05, **< 0.01). (C) Kaplan–Meier analysis of overall survival in CRC patients according to ZNF217 expression level. The ZNF217 positive group (n = 55) showed significantly shorter survival compared with the negative group (n = 27; P = 0.0405: log-rank test).
Table 1.
Association between patients, characteristics and ZNF217 expression in 82 CRC cases.
Figure 2.
Effects of ZNF217 on proliferation, migration and invasion of SW480 cell lines (A) Expression of ZNF217 in CRC cell lines.
(B) Reduction of ZNF217 expression by transfecting siRNA-ZNF217 significantly inhibited proliferation (* P < 0.05) and (C) migration and invasion of SW480 cells (200× magnification, * P < 0.05) in comparison with parental and negative controls.
Figure 3.
MiR-203 negatively regulates ZNF217 expression via the ZNF217 3’-UTR.
(A) The putative miR-203 binding sequences in ZNF217 3’-UTR. (B) Luciferase activity assay was performed for the HEK293T cells cotransfected with pmiR-REPORTTM vectors containing WT-ZNF217 3’-UTR or MUT-ZNF217 3’-UTR sequences and miR-203 mimics. Data are presented as normalized fold change in luciferase activity. (C, D) ZNF217 mRNA and protein were determined in SW480 cells transfected with miR-203 mimics or miR-negative control by qRT-PCR and Western blot, respectively. (E) Inverse correlation between ZNF217 mRNA expression and miR-203 levels in CRC tissues was analyzed using Pearson’s correlation analysis.
Figure 4.
Effects of miR-203 on proliferation, migration and invasion of SW480 cell lines.
(A) Ectopic expression of miR-203 by transfecting miR-203 mimics significantly reduced proliferation of SW480 cells, in comparison with parental and negative controls (* P < 0.05). (C) Ectopic expression of miR-203 notably inhibited cell migration and invasion of SW480 cells (200×magnification, * P < 0.05). Inversely, inhibition of miR-203 expression by transfecting miR-203 inhibitors simultaneously (B) promoted proliferation (*P < 0.05) and (D) migration and invasion of SW480 cells, compared with parental and negative controls (200×magnification, *P < 0.05). Figure is a representative of 3 experiments with similar results.
Figure 5.
Functional effects of ZNF217 downregulation and miR-203 upregulation on SW480 cells.
(A) Effective suppression of ZNF217 protein expression by ZNF217 siRNA and miR-203 mimics respectively and combinedly. Note that ZNF217 expression is more efficiently suppressed by combined treatment. Suppression of ZNF217 simultaneously resulted in (B) significant inhibition of cell growth and (C) migration and invasion (200×magnification) of SW480 cells compared with negative controls. Note the synergistic inhibitory effect of combination of ZNF217 siRNA and miR-203 mimics, compared with either of them alone (*P < 0.05).