Tissue factor (TF) is known to play a role in tumor progression. In retinoblastoma, the expression and role of TF has not been determined yet. Herein, we demonstrated for the first time that TF is closely related to the proliferation of retinoblastoma cells, which could be therefore effectively suppressed by blockade of the TF pathway. TF was selectively expressed on the areas of highly mitogenic activity in an orthotopic transplantation mouse model of retinoblastoma. In addition, the levels of TF expression in retinoblastoma cells were elevated after FGF2 treatment, whereas the proliferative effect of FGF2 on retinoblastoma cells was significantly inhibited by blockade of the TF pathway via TF pathway inhibitor (TFPI). Interestingly, retinoblastoma cells cultured with FGF2 showed increased phosphorylation of both Akt and ERK1/2. Addition of TFPI nearly abolished the FGF2-induced phosphorylation of Akt and ERK1/2 in retinoblastoma cells. Therefore, our data suggest that TF expression in retinoblastoma cells is closely related to tumor cell proliferation and TFPI has the potential to inhibit retinoblastoma cell proliferation via the inhibition of both Akt and ERK1/2 activation.