Effect Of Hedgehog Signaling Pathway Inhibitor On Biological Behavior Of Intrahepatic Cholangiocarcinoma Cell Line RBE

Objective:To investigate the effect of cyclopamine, a Hedgehog(Hh) signaling pathway inhibitor, on the biological behaviour of intrahepatic cholangiocarcinoma cell line RBE.Methods:The proliferation of RBE cells was detected by Cell counting with typan blue dyeing and MTT assay. The apoptosis rate was analyzed by the flow cytometry. Transwell invasive cabin assay was used to detect the invasion ability and Western blotting was used to determine the protein expression of Gli1 and MMP-9 in the RBE cells before and after cyclopamine treatment.Results:1. Cyclopamine inhibited the growth of RBE cells in time and dose dependent manners.2. After cyclopamine treatment for 24 h, 48 h, 72 h, the apoptotic rates were significantly higher than those in control group.3. In control group, the number of cells invading through the Matrigel of invasion chamber was(154.52±13.61), while in experimental group it was(62.00±12.17), indicating that the invasion ability of the cells declined significantly.4. Western blotting showed that the protein levels of Glil and MMP-9 in the RBE cells were decreased after treatment with cyclopamine for 24 h and 48 h.Conclusion:1. The Hh signaling pathway inhititor cyclopamine suppresses the proliferation of RBE cells.2. The Hh signaling pathway inhititor cyclopamine promotes the apoptosis of RBE cells.3. The Hh signaling pathway inhititor cyclopamine inhibits the invasion ability of RBE cells.In a word, the activity of Hh signaing pathways is possible relative with the occurrence, development, metastasis and prognosis of ICC.