Objective To investigate the expression and significance of interleukin-4 (IL-4)/triggering receptor expressed on myeloid cells-2 (TREM-2) signaling pathway-related factors in the tissues of mouse external auditory canal cholesteatoma (EACC). Methods Fifteen C57BL6/J mice were randomly assigned to an experimental group (n=11) and a control group (n=4). EACC model was established in the experimental group via external auditory canal ligation. Three months after the surgery, the external auditory canal tissues were collected from the mice of the experimental and the control group for real-time quantitative polymerase chain reaction, Western blot and enzyme-linked immunosorbent assay to detect the mRNA and protein expression levels of IL-4, TREM-2, IL-6, IL-8, IL-1β and tumor necrosis factor-α(TNF-α). Results The modeling success rate in the experimental group 3 months after surgery was 91% (10/11). Compared with the control group, the mRNA and protein expression levels of TREM-2, IL-6, IL-8, IL-1β, and TNF-αin the EACC tissues of the experimental group were significantly increased (P<0.05), while the expression level of IL-4 was markedly reduced (P<0.05). Conclusion The IL-4/TREM-2 signaling pathway and the inflammatory response it mediates may be involved in the occurrence and development of EACC in mice.