High-throughput mRNA-sequence of MCM7 and SHCBP1 knock down hepatocytes with IL-1ß stimulation
Liver fibrosis, characterized by the excessive deposition of extracellular matrix, remains a formidable challenge due to its complex and poorly understood mechanisms. In this study, we aimed to elucidate the role of Minichromosome Maintenance Complex Component 7 (MCM7) and SHC SH2-Domain Binding Protein 1 (SHCBP1) in liver fibrosis, focusing on their regulatory network and interaction. Stable hepatocyte cell lines with knockdown of MCM7 and SHCBP1 were constructed and subsequently stimulated with IL-1ß to mimic inflammatory conditions. RNA sequencing was conducted to identify differentially expressed genes and pathways affected by the knockdown. Our analysis revealed significant changes in gene expression profiles, particularly in pathways related to fibrosis and inflammation. We discovered that MCM7 interacts with SHCBP1, which in turn binds to RACGAP1. This interaction activates the SHCBP1-RACGAP1-STAT3 signaling axis, leading to increased IL11 expression. Elevated IL11 levels contribute to the activation of hepatic stellate cells and the progression of liver fibrosis. The findings demonstrate that MCM7 is an important regulatory factor in liver fibrosis and promotes IL11 expression through the SHCBP1-RACGAP1-STAT3 signaling pathway, driving hepatic stellate cell (HSC) activation and fibrosis progression. Overall, our findings highlight a mechanistic pathway where MCM7 and SHCBP1 collaborate to regulate IL11 expression, thereby influencing liver fibrosis progression. Overall design: To investigate the cooperative function of MCM7 and SHCBP1 in hepatocytes with IL-1ß stimulation, we established cell lines in which each target gene has been knocked down by shRNA. We then performed gene expression profiling analysis using data obtained from RNA-seq of 3 different cell lines with or without IL-1ß stimulation. Comparative gene expression profiling analysis of RNA-seq data for hepatocytes and their KD derivatives (shMCM7 and shSHCBP1) under IL-1ß stimulation was conducted to determine the impact of these knockdowns