Fig 1: Cancer cell-derived MAMDC2 promotes MYLK expression in CAFs. (A) Expression of MAMDC2 and MYLK in CRC with different TSRs. Original magnifications 40× and 100×. n (SP-L) = 85. n (SP-H) = 36. (B) Predicted subcellular localization and membrane association of MAMDC2. (C) Western blot analysis of MAMDC2 in NCM460, HCT116, and RKO culture supernatant. (D) Western blot analysis of MAMDC2 overexpression in RKO and its regulatory effect on MYLK expression in both RKO (left) and CAFs (right).
Fig 2: Co-expression patterns of MAMDC2 and MYLK in colorectal cancer. (A) Correlation between MAMDC2 and the top 10 genes in the EMT gene set and myogenesis gene set. (B) Correlation between MAMDC2 and MYLK. (C) The expression of MYLK in CRC tissues. n (normal) = 359. n (tumor) = 383. (D) The expression level of MYLK in different CMS subtypes of CRC. n (CMS1) = 84. n (CMS2) = 145. n (CMS3) = 71. n (CMS4) = 138. (E) Localization and expression of MAMDC2 and MYLK in single-cell sequencing data (GSE166555 and EMTAB8107). * p < 0.05, **** p < 0.0001.
Fig 3: MAMDC2 regulates epithelial–mesenchymal transition in colorectal cancer. (A) MAMDC2 single gene difference analysis. (B) GO and KEGG based on MAMDC2 single gene differential expression. (C,D) EMT gene set and myogenesis gene set from GSEA based on MAMDC2 single gene differential expression gene set. (E) Correlation between MAMDC2 and EMT-related genes. (F) The real-time RT-PCR analysis of CDH2, MMP2, MMP9, and SNAI1 with MAMDC2 overexpression (above) or knockdown (below). (G) Western blot analysis of EMT-related proteins (N-cadherin, MMP2, and Snail) in RKO with MAMDC2 overexpression (left) and HCT116 with MAMDC2 knockdown (right).
Fig 4: MAMDC2 expression patterns in primary and metastatic colorectal cancer. (A) MAMDC2 expression in peritoneal and liver metastases from the GSE183916 and GSE245552 datasets. (B) Representative images (above) and scores (below) of immunohistochemical analysis of the expression of MAMDC2 in primary and metastatic lesions of colorectal cancer. Original magnifications 40× and 100×. n (peritoneal metastases) = 22. n (liver metastases) = 18. n (lymph node metastases) = 26. * p < 0.05, ** p < 0.01, **** p < 0.0001, ns = not significant.
Fig 5: Differential expression and survival analysis of MAM domain-containing genes in colorectal cancer. (A) Identification of 18 MAM domain-containing genes via the InterPro database. (B) Volcano plot of differential mRNA expression of MAM domain-containing genes between normal and CRC tissues. (C–F) The Kaplan–Meier Plotter was used to determine the correlation between the mRNA levels of MAMDC2 (C), MEP1A (D), MEP1B (E), and EGFL6 (F) and the survival of CRC patients. p < 0.05.
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