Objective: To investigate the expression of FIG-ROS fusion gene in intrahepatic cholangiocarcinoma (ICC) cells and the effects of its intervention on biological behavior of ICC cells. Methods: ROS protein expression in 4 different specimens of ICC tissue and 3 types of ICC cell line (HUCCT1, REB and QBC939) was determined by Western blot analysis; the ROS positive cell line was selected for use and after transfection with a series of plasmids containing different sequences of ROS-shRNAs or FIG-shRNAs respectively, the protein expressions of ROS and FIG in the cells were measured by Western blot analysis. The sequences of ROS-shRNA and FIG-shRNA with highest inhibitory effect on ROS and FIG expression were chosen, which were alone or in combination transfected into the above cells, and after that, the cell proliferation, apoptosis, cell cycle and colony formation were observed. Results: Two specimens of ICC tissue and one ICC cell line (HUCCT1) showed positive ROS expression. Transfection of ROS1-6290 shRNA and FIG-363 shRNA had the most remarkable inhibitory effect on ROS and FIG expression, respectively. Compared with the HUCCT1 cells without any transfection, lone FIG- 363 shRNA transfection had no obvious effect on proliferation, apoptosis or cell cycle phase (all P>0.05), but significantly reduced the colony formation of the cells (P<0.05); either ROS1-6290 shRNA transfection alone or in combination with FIG-363 shRNA showed significant effects of suppression of proliferation, induction of apoptosis and cell cycle arrest and inhibition of colony formation, and these effects were more remarkable in cells with combined transfection (all P<0.05). Conclusion: Some kinds of ICC have FIG-ROS fusion gene expression, and the combined inhibition of the two genes may probably provide a hopeful targeted treatment approach for these ICC.