BMI1 and cholangiocarcinoma: Next, to examine the effect of Bmi1 overexpression in AY-CCA, we injected pT3-EF1ɑ-GFP or pT3-EF1ɑ-BMI1-V5 plasmids along with AY plasmids and evaluated tumorigenesis at 6 to 8 weeks after HDTVI (Fig. 4B), and successful Bmi1 upregulation was confirmed by qPCR (Fig. 4G).