Fig. 1
From: C1Q+ TPP1+ macrophages promote colon cancer progression through SETD8-driven p53 methylation
SETD8 is highly expressed in CRC cells and p53K382me1 is an independent prognostic factor associated with poor prognosis in CRC patients. A Flow chart of the methodological approach to select chromatin modifying enzymes with positive fold change and p-value < 0.05 overexpressed in tumors derived from CRC patients compared with healthy subjects (TCGA dataset) and enriched in CRC cells compared to normal enterocytes (scRNA-seq data from GEO, GSE 200997). The top 15 significantly enriched pathways were obtained by Enrichment Pathway Analysis based on the Reactome Gene set (Molecular Signature Database, MSigDB), utilizing msigdbR library and p < 0.05. Pathways related to p53 signaling are among the top significantly enriched pathways. Venn Diagram showing the common genes enriched in the top 5 p53-related pathways. B Relapse-free survival (RFS) analysis of TP53 expression in CRC patients from R2 database (Tumor colon, Marisa dataset). Statistical significance was calculated using the log-rank test. C TCGA analysis of KMT5A (SETD8) relative mRNA expression levels in normal and tumor tissue derived from CRC patients (COAD dataset). D Immunohistochemical analysis of SETD8 and p53K382me1 on healthy colon mucosa of a healthy subject and on tumor tissue of a p53WT CRC patient. Scale bars, 40 µm. E Immunoblot analysis of the indicated total and histone proteins in SY5Y NB cells, HCT116 and LOVO colon cancer cells, and 11 CR-CSphCs (left panel). Densitometry analysis of p53K382me1 protein levels compared with p53 total protein levels measured as relative density units (RDU) (right panel). F Immunoblot analysis of the indicated proteins in CRL-1790 and CRL-1831 healthy colon cells, in HCT116 colon cancer cells and in CR-CSphC#8. β-actin was used as loading control. G Immunofluorescence analysis of p53K382me1 and CD44v6 in tumor tissue derived from a p53WT CRC patient. Nuclei were counterstained by Toto-3. Scale bars, 40 µm. H Immunohistochemical analysis of CD44v6 and immunofluorescence analysis of p53K382me1 and SETD8 on tumor tissue of two CRC TMAs representative of high staining intensity (#1) or lack of staining (#2), respectively. Nuclei were counterstained by Toto-3. Scale bars, 40 µm. I Immunofluorescence analysis of p53K382me1 in CRC TMA patients in tumor tissue (T) or in the immune cell infiltration (I). Four different conditions are indicated (Tum + /Inf-, Tum + /Inf + , Tum-/Inf-, Tum-/Inf +). Nuclei were counterstained by Toto-3. Scale bars, 40 µm. J Probability of survival in CRC TMA patients based on the p53K382me1 expression in tumor tissue (Tum ±) and/or in the immune cell infiltration (Inf ±). K Disease-free survival probability in CRC patients based on p53K382me1 expression in tumor tissue or in the immune cell infiltration. See also Tables S1, Table S2, Table S3 and Figure S1