Deciphers the role of myofibroblasts in microvascular invasion of hepatocellular carcinoma

Xiang LiuJiangyang PanLijia WangQiao XieTianbo FanQi Wang^*

• Department of Radiology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, China

Hepatocellular carcinoma (HCC) often exhibits microvascular invasion (MVI), a feature with unclear mechanisms. Therefore, it is crucial to resolve its related cellular populations and molecular networks using single-cell analysis.Both single cell RNA sequencing (scRNA-seq) and RNA-seq data for HCC were obtained from public databases. scRNA-seq data were clustered and annotated using Seurat. DAVID, CellChat and Monocle 2 were used for scRNA-seq functional enrichment analysis, intercellular communication analysis and cell trajectory analysis, respectively. We further assessed key gene expression in HCC cell lines and examined their effects on cell functions using CCK-8, scratch, and transwell assays.The ecosystem consisted of myofibroblasts (MFs), hepatocytes, proliferative hepatocytes, endothelial cells, dendritic cells, proliferative NK/T cells, plasma B cells and macrophages in HCC was revealed. MFs showed the greatest difference between MVI absent and present patients and were subdivided into five clusters. Key genes for angiogenesis are overexpressed in MF2 and enriched in the pathways of angiogenesis, cell migration, cell proliferation, and signal transduction. Pseudotime analysis revealed MF2 from MVI-present patients clustered at the terminal state and positively correlated with angiogenesis. CAMK2N1 in the markers of MF2 was significantly associated with later M stage and poor prognosis. Further cellular assays showed that CAMK2N1 expression was down-regulated in HCC cells and its knockdown increased the proliferation, migration and invasion levels of cancer cells.This study highlighted the role and potential mechanism of MFs in promoting MVI formation and provides a potential marker for HCC prognosis among MFs markers.This study aimed to identify and characterize the major cell populations and potential molecular regulatory networks that 删除[Administrator]:HCC as a multi-vascular malignancy often exhibits histological features of MVI.