In this work, we show that PTOV1 promotes the invasion and anchorage-independent growth of prostate cancer cells while it acts as a novel repressor of the Notch target genes HES1 and HEY1. Reciprocally, a constitutively activated Notch1 receptor decreases anchorage-independent growth and invasion in vitro. Here, HEY1 is linked to Familial prostate cancer.