Background: Cholangiocarcinoma (CCA) is one of the most prevalent diseases in the South Asia. Failing to determine this disease on time tends to be a great challenge in its diagnosis. In several human malignancies, indirubin-3'-oxime (I3O) has been found to decrease cell growth, cause cessation of cell cycle, and cell death. Objective: The anticancer effects of I3O on human CCA cells will be demonstrated by exhibiting induced cessation of cell cycle and apoptosis in I3O-treated cells. Methods: The viability, apoptosis of cells and cessation of cell cycle were all measured. Analysis using Western blotting, flow cytometry and immunofluorescence. Results: According to the findings, in CCA cells, I3O treatment inhibits cell growth and induces cessation of cell cycle as well as caspase-dependent death. Conclusion: These data imply that I3O, by regulating the cell cycle and generating apoptosis, could inhibit tumour growth and hence be a viable treatment drug for human CCA.