In conclusion, it was verified by in vivo experiments that pGCsilencer-U6/Neo/GFP could inhibit the proliferation, migration, and invasion of cholangiocarcinoma, while promoting the apoptosis of tumor cells, whose mechanism may be related to the ability of VEGF silencing to enhance apoptosis and inhibit the expressions of MMP2 and MMP9. Here, MMP9 is linked to cholangiocarcinoma.