GLI3 Promotes Invasion and Predicts Poor Prognosis in Colorectal Cancer

Figure 3

GLI3 promotes EMT via regulating p-ERK signaling pathway. (a) TCGA CRC samples were divided into two groups: epithelial-like phenotype (green) and mesenchymal-like phenotype (red). (b) GLI3 expression was significantly elevated in the mesenchymal-like group. (c) Correlation heat map indicated that GLI3 was negatively correlated with CDH1, and positively correlated with TWIST1, CDH2, ZEB1, ZEB2, and VIM. (d) Details of the correlation between logTPM-transformed GLI3 and CDH2, TWIST1, VIM, and ZEB1. (e–g) Two CRC cell lines DLD1 and SW480 were used to knockdown GLI3 expression. In both cell lines, specific siRNA targeting GLI3 had an optimal efficiency of silencing, and western blot was further performed to validate the knockdown efficiency at the protein level. qRT-PCR and western blot analyses demonstrated that the EMT markers were significantly downregulated in GLI3-silencing cells at both mRNA level and protein level. (h, i) Silencing of GLI3 greatly attenuated both the migratory and invasive capacities of DLD1 and SW480 cells in response to siRNA-treatment. (j) GSEA analysis indicated that high expression of GLI3 is involved in the function of positive regulation of ERK1/2 cascade. (k) Western blot demonstrated that the pERK1/2 activation was significantly attenuated in GLI3-knockdown groups.

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