Objective To explore the effect of ras-related C3 botulinum toxin substrate 3 (RAC3) on the proliferation, migration and invasion abilities of early trophoblast cells. Methods Villius samples from 20 unexplained spontaneous abortion (SA) and 20 induced abortion (IA) patients were collected between May 2015 and May 2016 in Changzheng Hospital of Navy Medical University (Second Military Medical University). qPCR was used to detect the expression of Rac3 mRNA in the villus tissues. mRNA Chip detection was performed on the placental tissues of 6.5, 14.5 and 19.5 days in mice. After interfering or overexpressing RAC3 in early human trophoblast cell line HTR-8/SVneo, the proliferation, migration and invasion abilities were detected by CCK-8 and Transwell assay, respectively. Results The expression of Rac3 mRNA was significantly lower in the villus tissue of unexplained SA patients than that in the villus tissue of IA patients (P<0.05). Rac3 mRNA expressions were significantly higher in the placental tissues of 6.5 and 14.5 days in mice than that in the placental tissues of 19.5 days (both P<0.05). Compared with the control group, the proliferation, migration and invasion abilities of the HTR-8/SVneo cells were significantly reduced by interfering RAC3 expression (all P<0.05), and the proliferation, migration and invasion abilities were significantly enhanced by overexpressing RAC3 (all P<0.01). Conclusion RAC3 plays an important role in regulating of the proliferation, migration and invasion of early trophoblast cells.