Transcriptome Sequencing Analysis of Human Colorectal Cancer Cell Lines After ECM1 Knockdown

To investigate the regulatory role of ECM1 in CCL20 expression in colorectal cancer, this study employed the human colorectal cancer SW620 cell line. Stable knockdown of ECM1 gene expression was achieved via shRNA, and single-cell transcriptome sequencing technology was used to systematically compare the transcriptomic differences between the ECM1 knockdown group (SW620-shECM1) and the negative control group (SW620-shNC). Sequencing analysis revealed that ECM1 knockdown significantly affected multiple signaling pathways, among which the TNF-a signaling pathway was notably activated, and the expression and activity of the key transcription factor JUNB underwent significant changes. Further mechanistic studies demonstrated that ECM1 regulates the transcription and expression of the downstream chemokine CCL20 by modulating the TNF-a/JUNB signaling axis. This study is the first to reveal the molecular mechanism by which ECM1 regulates CCL20 through the TNF-a-JUNB pathway in colorectal cancer at the single-cell level, providing a new perspective for understanding the role of ECM1 in the tumor microenvironment and immune regulation. Overall design: The human colorectal cancer SW620 cell line was used to explore ECM1's regulatory role in CCL20 expression. Stable ECM1 knockdown was established in SW620 cells using shRNA (SW620-shECM1 group), with a negative control group (SW620-shNC) set simultaneously. Single-cell transcriptome sequencing was performed to compare transcriptomic differences between the two groups, focusing on analyzing changes in signaling pathways and key molecule expression related to ECM1-mediated CCL20 regulation.