MicroRNAs (miRNAs) encoded by latency-associated transcript are associated with both latent and acute stages of herpes simplex virus 2 (HSV-2) infection.In this study,miRNA-H4-5p and miRNA-H4-3p were ectopically expressed in HeLa cells to explore potential cellular targets of viral miRNAs and demonstrate their potential biological functions.The results showed that miRNA-H4-5p could reverse apoptosis induced by actinomycin D (Act-D) and promote cell cycle progression,but miRNA-H4-3p had no such obvious functions.Bioinformatics analysis,luciferase report assay,quantitative reverse transcription polymerase chain reaction (qRT-PCR),and Western blotting demonstrated that miRNA-H4-5p could bind to the 3\'-untranslated region (UTR) of cyclin-dependent kinase inhibitor 2A (CDKN2A) and cyclin-dependent kinase-like 2 (CDKL2) to negatively regulate their expression.We verified that these two targeted genes were associated with cell apoptosis and cell cycle.Furthermore,in HeLa cells infected with HSV-2,we detected significantly reduced expression of CDKN2A and CDKL2 and demonstrated the negative regulation effect of miRNA-H4-5p on these two target genes.Our findings show that viral miRNAs play a vital role in regulating the expression of the host\'s cellular genes that participate in cell apoptosis and progression to reshape the cellular environment in response to HSV-2 infection,providing further information on the roles of encoded herpesvirus miRNAs in pathogen-host interaction.