Single-cell sequencing analysis of human colorectal cancer tissues to explore the tumor microenvironment and cell heterogeneity
In this study, single-cell RNA sequencing via the 10x Genomics Chromium platform was conducted on fresh surgical colorectal cancer tissues from two patients, one with liver metastasis (advanced stage) and one without distant metastasis (early stage). After dissociating tissues into single-cell suspensions and sequencing, raw data underwent quality control, alignment, and gene expression quantification. Through differential expression and clustering analyses, we compared gene expression profiles and cell compositions between the two groups. Our aim was to identify unique cell populations, their characteristic gene expression patterns, and genes differentially expressed between advanced and early stages. These findings may offer new insights into colorectal cancer progression mechanisms and aid in developing more effective stage-specific diagnostic, treatment, and prognostic strategies. Overall design: Fresh surgical resection tissues from two colorectal cancer patients were collected, one with liver metastasis (advanced stage) and the other without distant metastasis (early stage). Tissues were dissociated into single-cell suspensions and subjected to single-cell RNA sequencing using the 10x Genomics Chromium platform. Cells were captured, barcoded, and sequenced.  Afterward, raw data underwent standard bioinformatics processing, including quality control, alignment, and gene expression quantification. Differential expression analysis was performed to compare gene expression between the two samples, aiming to identify genes differentially expressed in advanced and early stages. Clustering analysis grouped cells by gene expression patterns to distinguish cell types and find stage-related differences. Overall, we aimed to uncover single-cell level differences between advanced and early colorectal cancer.