Due to its high incidence and mortality rates, colorectal cancer (CRC) has become the focus of research. Vitamin D has anticancer functions in a variety of cancers, but the relationship between vitamin D status and CRC risk and survival is inconclusive. The purpose of this study was to explore the effect of vitamin D on the proliferation and invasion of CRC cells to find an effective way to treat CRC. The results showed that vitamin D could inhibit SW480 cell progression in vitro. The results showed that vitamin D significantly blocked the Notch1 pathways by decreasing NOTCH1 protein expression. In addition, we found that Notch1 overexpression could significantly reverse the inhibitory effect of vitamin D on SW480 cells. These results reveal novel insights into the mechanisms of vitamin D anticancer activity in CRC development and progression.