Objective To explore the molecular mechanism by which dendritic cells (DC) sense changes in the extracellular mechanical microenvironment and dynamically adjust their migration behavior. Methods Hydrogel substrates with varied stiffness were constructed to investigate the influence of the mechanical microenvironment on DC migration behavior. Fbrotic rat liver model was established, in combination with immunohistochemistry experiments, to investigate the effect of liver fibrosis on DC migration capability. Furthermore, potential key molecules involved in the mechanical sensing cascade during DC migration were analyzed through single-cell sequencing data from human/mouse fibrotic livers, and RT-qPCR was used to examine the expression levels of the above key molecules in mouse DCs on substrates of different stiffness. Results The migration capability of DC on stiff substrates was significantly lower than that on soft substrates; DC infiltration in fibrotic rat livers increased; 682 differentially expressed genes (DEGs) were found between liver-infiltrating DCs from cirrhosis patients and normal people. Further focusing on genes relevant to cytoskeleton regulation and migration based on KEGG and GO pathway enrichment analysis, 12 potential key molecules mediating the stiffness sensing during DC migration were screened out. Among them, the expression levels of AIF1, GPR65, MYL12B, RAC1, and RHOG genes were up-regulated in patients with liver cirrhosis, while ACTB, ACTG1, ARF6, CDC42, COTL1, PFN1, and TMSB10 were down-regulated. Subsequently, the expression levels of ACTB and CDC42 were down-regulated in mouse DC on stiff substrates, which was consistent with the circumstance of liver-infiltrating DC in human cirrhotic patients. Conclusion The fibrosis in liver potentially induces impaired DC migration, resulting in increased DC infiltration. ACTB and CDC42, are promising regulators that mediate DC stiffness sensing during DC migration. In this paper, we preliminarily explore the potential key molecules that enable DCs to sense the alterations of environmental stiffness and dynamically adjust their migration behavior. Interventions based on the above molecules would potentially modulate DC migration efficiency in vivo, providing theoretical basis and inspiring novel strategies for optimizing DC mediated anti-tumor immune functions.