Fig 2: CMTR1-regulated tumor cell proliferation and immune responses are STAT3-dependent.A–G Changes of STAT3 knockdown on CMTR1-induced inflammatory mediators. H–J Effect of STAT3 knockdown on CMTR1-induced tumor cell proliferation, as assessed by CCK8, colony formation, and EdU incorporation assays (scale bar, 50 µm). *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 3: CMTR1 regulates the expression of a group of genes involved in the cell cycle.A RNA-seq was conducted on RKO cells transfected with control siRNA (si-Ctrl) or CMTR1 siRNA (si-CMTR1). A pie chart was used to show genes positively and negatively regulated by CMTR1. B, C The expression levels of genes positively and negatively regulated by CMTR1 are displayed in a heatmap and a box plot. D Hallmark gene set analysis revealed the gene term with the greatest enrichment of genes positively regulated by CMTR1. E–G Graphical presentation of the effects of CMTR1 on representative cell cycle-related genes, such as CDKN1A, CDK6, and CCND1, as determined by RNA-seq. H–Q Expression of CDKN1A, CDK6, and CCND1, measured by RT?qPCR and western blotting in CMTR1 knockdown and CMTR1-overexpressing cells. *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 4: CMTR1 promotes colorectal cancer cell growth both in vitro and in vivo.A, B CMTR1 mRNA expression was measured by RT?qPCR in CMTR1 knockdown and CMTR1-overexpressing cells. C–H CCK8, colony formation, and EdU incorporation assays were performed to evaluate cell proliferation (scale bar, 50 µm). I Subcutaneous tumorigenesis in mice was evaluated after CMTR1 knockdown. *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 5: CMTR1 affects the expression of inflammatory factors in colorectal cancer cells by transcriptionally regulating STAT3.A KEGG pathway analysis revealed that genes differentially regulated by CMTR1 knockdown were significantly enriched in the JAK-STAT signaling pathway. B, C STAT3 mRNA expression was measured by RT?qPCR in CMTR1 knockdown and CMTR1-overexpressing cells. D STAT3 and pSTAT3 protein expression levels were assessed by western blotting after knockdown and overexpression of CMTR1. E Correlation analysis of CMTR1 and STAT3 expression using public TCGA transcriptome datasets via the gene expression profiling interactive analysis (GEPIA) server. F, G RKO cells were subjected to ChIP analysis with control IgG or an anti-CMTR1 antibody, followed by RT?qPCR analysis using two primer sets (P1 and P2) targeting the two predicted CMTR1 binding sites in the STAT3 promoter region. H, I After knockdown of CMTR1, RKO cells were subjected to ChIP analysis with control IgG or an anti-RNAPII antibody, followed by RT?qPC analysis using two primer sets (P1 and P2) targeting the two predicted RNAPII binding sites in the STAT3 promoter region. J–Q Relative mRNA levels of inflammatory mediators after knockdown and overexpression of CMTR1. *P < 0.05, **P < 0.01, and ***P < 0.001.