Abstract: Objective To analyze the changes and value of protein expression profile following overexpression of human cytomegalovirus (HCMV ) -encoded miR-US25- 1-3p in human mononuclear cell line( THP-1). Methods hemv-miR-US25- 1-3p mimics were transfected into THP-1 cells using liposomes to construct hemv -miR-US25-1-3p overexpressed cell line. The transfection effects were verified by RT-qPCR. The quantitative analysis for proteomics of THP-1 cell line after hemv-miR-US25-1-3p overexpression was performed by using tandem mass spectrometry labeling combined with liquid chromatography tandem mass spectrometry. The physiological and pathological functions of diferentially expressed proteins were analyzed by bioinformatics. The expression changes of the important differential proteins were verified by western blot. Results Compared with the controls, the hemv-miR-US25-1-3p overexpressed THP-1 cells showed differential expressions of 65 types of proteins, of which 17 were up-regulated and 48 were down-regulated. The results of annotation and functional prediction by GO, COG, and KEGG databases indicated that differentially expressed proteins were mainly involved in inflammatory response signaling pathways，metabolic processes, cellular and genetic information functions and associated with multiple diseases, e.g., cancer, viral myocarditis , nonalcoholic fatty liver disease, primary immunodeficiency disease, rheumatoid arthritis ,multiple pathogen infections, Alzheimer' s disease, Huntington's disease and others. The results of literature research and bioinformatics analysis found that tumor necrosis factor related apoptosis inducing ligand ( TRAIL) and activated regulatory normal T cell expression and secretion factors ( RANTES ) involved in antiviral and inflammatory responses among the 48 types of down-regulated proteins. The down-regulated expressions of the above proteins were verified by western blot. Conclusion HCMV encoded hemv-miR-US25-1-3p in THP-1 cells could affect the host protein expression profile, and may promote virus latency and escape from host cell killing by inhibi- ting the expression of TRAIL and RANTES.