IL-6 induced EMT through STAT3 phosphorylation.

(A) Western blot of HCT 116 and COLO 205 cells treated with 50 ng/mL IL-6. (B) Western blot analysis of HCT 116 and COLO 205 cells after exposure to increasing concentrations of IL-6. Fold-, fold-change; pSTAT3, phosphorylated STAT3; β-actin, beta actin.</p

Effect of metformin on IL-6-induced STAT3 phosphorylation and <i>MMP9</i> expression in the human colon cancer cell line COLO 205.

(A) Western blot analysis of metformin- and IL-6- treated colon cancer cells (HCT 116). (B) Western blot analysis of metformin- and siRNA STAT3 (siSTAT3)—treated colon cancer cells (HCT 116) (C) qPCR analysis of metformin- and IL-6-treated colon cancer cells. Fold-, fold-change; Met, metformin. *p p < 0.01.</p

IL-6 induced EMT in colon cancer cells.

(A) Cell viability assay for colon cancer cell lines. (B) Migration assay for colon cancer cell lines treated with different concentrations of IL-6 for the indicated times. (C) Morphological changes in colon cancer cells upon treatment with IL-6 at the indicated concentrations. Scale bar = 300 μm, HCT 116. *p p < 0.01.</p

Pathway analysis of TCGA patients.

(A) Canonical pathways in TCGA colon cancer patients as assessed by IPA. IL-6 and colorectal metastasis signaling were decreased. (B) GSEA showed that EMT and IL-6 signaling were reduced in the metformin-predicted group significantly. (C) mRNA expression of IL6, FN1, and MMP9 was decreased in the metformin-predicted group (TCGA dataset). *p p < 0.01, JAK: Janus kinase, ES: enrichment score, FDR: false discovery rate.</p

Metformin inhibited IL-6-induced EMT in the human colon cancer cell line.

(A) Migration assay for colon cancer cells incubated with IL-6 and metformin (HCT116). (B) Invasion assay for colon cancer cells (HCT 116). (C) Immunofluorescence confocal microscopy analysis of HCT 116 colon cancer cells treated with IL-6 and metformin. Met, metformin. *p p < 0.01.</p

Gene expression profiles of colon cancer samples stratified by the metformin-treated gene signature of colon cancer cells (GSE67342).

(A) Heatmap of the gene signature from the metformin-treated LoVo colon cancer cells and the control LoVo colon cancer cells (left). The prediction and grouping of TCGA colon cancer patients by the signature from the LoVo cells (right). (B) Kaplan–Meier survival plots of TCGA colon cancer patients by the results of prediction. The metformin-predicted group showed the better survival rate.</p

Kaplan–Meier Plots of DFS rates of all patients grouped by AJCC stage.

<p>Patients were stratified by risk level according to the five predictors (A to E). <i>P</i> values are based on the log-rank test.</p

CONSORT flow diagram for selection of prediction models.

<p>CONSORT flow diagram for selection of prediction models.</p

Kaplan–Meier survival plots of the DFS rates of VI patients stratified by risk level according to the five genomic predictors (A to E).

<p><i>P</i> values are based on the log-rank test. Int, intermediate.</p

Multivariate Cox proportional hazard regression analyses of DFS with clinical variables and genomic predictors.

<p>Multivariate Cox proportional hazard regression analyses of DFS with clinical variables and genomic predictors.</p