Objective  To investigate the effects of Notch2 and Notch3 on the migration and invasiveness of trophoblast cell lines BeWo and JAR. Methods  The changes of migration and invasiveness of two trophoblast cell lines were observed by over-expression and intervention of Notch2 and Notch3 of BeWo and JAR cells. Results  The migration of BeWo cells at 48 h significantly decreased after the intervention of Notch2, while the migration at 72 h and invasiveness after the intervention of Notch2 did not change remarkably. The migration of BeWo cells at 48 h significantly increased after the over-expression of Notch3, while significantly decreased at 72 h. The invasiveness at 48 and 72 h significantly increased after the over-expression of Notch3. The migration of JAR cells at 48 and 72 h significantly decreased after the intervention of Notch3, while the invasiveness after the intervention of Notch3 and the migration and invasiveness after the over-expression of Notch2 didn’t change remarkably. Conclusion  Notch2 and Notch3 can affect the migration and invasiveness of trophoblast cell lines BeWo and JAR.