Downregulated extracellular matrix-associated CILP, MFAP4, and MMRN1 genes reveal links to immune infiltration and clinical outcomes in colorectal cancer: bioinformatics and experimental validation.
Colorectal cancer (CRC) is a leading cause of cancer mortality. The extracellular matrix (ECM) plays a critical role in tumor progression, and its associated genes may serve as prognostic biomarkers. This study investigated three ECM-related genes, CILP, MFAP4, and MMRN1, to evaluate their expression and clinical significance in CRC development.
RNA-seq data from TCGA-COAD and microarray datasets (GSE23878, GSE89076) were analyzed to identify differentially expressed genes (DEGs). Bioinformatics analyses included functional enrichment, protein-protein interaction (PPI) network construction, and immune infiltration evaluation. Experimental validation was performed using RT-qPCR on 20 paired CRC and adjacent non-cancerous tissues. Associations between gene expression, clinicopathological features, and overall survival were assessed.
Both RNA-seq and microarray analyses revealed significant downregulation of CILP, MFAP4, and MMRN1 in CRC tissues compared to non-cancerous samples (p < 0.05). RT-qPCR confirmed lower expression levels in tumor tissues (p < 0.0001). TIMER database analysis for immune cell infiltration revealed positive correlations between target genes and tumor-infiltrating lymphocytes, particularly CD8+ T cells and macrophages. Moreover, MFAP4 expression showed a significant association with lymph node status (p < 0.05), while both MFAP4 and MMRN1 were found to be correlated with smaller tumor size (p = 0.04). Survival analysis showed non-significant trends toward poorer prognosis in patients with lower gene expression.
These findings demonstrated that CILP, MFAP4, and MMRN1 function as potential tumor suppressors and immunomodulators in CRC. Their significant downregulation in tumor tissues, along with associations with immune infiltration and clinical features, suggest a possible role in CRC pathogenesis and make them potential therapeutic targets.
RNA-seq data from TCGA-COAD and microarray datasets (GSE23878, GSE89076) were analyzed to identify differentially expressed genes (DEGs). Bioinformatics analyses included functional enrichment, protein-protein interaction (PPI) network construction, and immune infiltration evaluation. Experimental validation was performed using RT-qPCR on 20 paired CRC and adjacent non-cancerous tissues. Associations between gene expression, clinicopathological features, and overall survival were assessed.
Both RNA-seq and microarray analyses revealed significant downregulation of CILP, MFAP4, and MMRN1 in CRC tissues compared to non-cancerous samples (p < 0.05). RT-qPCR confirmed lower expression levels in tumor tissues (p < 0.0001). TIMER database analysis for immune cell infiltration revealed positive correlations between target genes and tumor-infiltrating lymphocytes, particularly CD8+ T cells and macrophages. Moreover, MFAP4 expression showed a significant association with lymph node status (p < 0.05), while both MFAP4 and MMRN1 were found to be correlated with smaller tumor size (p = 0.04). Survival analysis showed non-significant trends toward poorer prognosis in patients with lower gene expression.
These findings demonstrated that CILP, MFAP4, and MMRN1 function as potential tumor suppressors and immunomodulators in CRC. Their significant downregulation in tumor tissues, along with associations with immune infiltration and clinical features, suggest a possible role in CRC pathogenesis and make them potential therapeutic targets.
Authors
Golestannejad Golestannejad, Kalantary-Charvadeh Kalantary-Charvadeh, Karimi Karimi, Khodadadi Khodadadi
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