Abstract: Objective　To explore the inhibitory effect of Newcastle disease virus (NDV) on the cell proliferation, migration and invasion by influencing the cell microfilament skeleton in the process of infecting cervical cancer HeLa cells. Methods　HeLa cells were divided into control group, NDV infection group (NDV F3 group) and cytochalasin D group. The morphological changes of the cells were observed through an optical inverted microscope. CCK-8,TUNEL and Hoechst 33258 staining were used to detect cell proliferation and apoptosis of cells. Scratch test and Transwell test were used to observe the effects of NDV F3 on the cell migration and invasion. Western blot assay was used to detect the expressions of F-actin, Ras homolog family member A (RhoA) and Rho kinase (ROCK)2 related microfilament frame work proteins after NDV F3 was applied to the cells. Results　After NDV F3 acting on the cells, the cells lost their normal morphology, and some cells were fused. As the infection time increased, the cells became round, fall off and rupture, leaving very few adherent cells.CCK-8 results showed that the cell proliferation rate decreased. TUNEL detection and Hoechst33258 staining showed that NDV F3 induced cell apoptosis, and the phenomenon of apoptosis was more significant when MOI=0.1. Western blot results showed that F-actin, RhoA and ROCK2 all had a downward trend after 36 h and 48 h (P＜0.01). In the scratch test at 24 h and 48 h, the migration rate and the migration rate were decreased in NDV F3 group and cytochalasin D group compared with those of the control group, and which was most significant at 48 h (P＜0.01).Compared with the control group, the number of cell invasions was significantly reduced in the  NDV F3 group and cytochalasin D group (P＜0.01). Conclusion　NDV F3 has inhibitory effects on the proliferation, invasion and migration of HeLa cells, and its mechanism may be related to the RhoA/ROCK signaling pathway related microfilament skeleton.