Effect of Hedgehog signaling pathway inhibitor on biological behavior of intrahepatic cholangiocarcinoma cell line RBE / 中国病理生理杂志

ABSTRACT

AIM:To investigate the effect of cyclopamine , a Hedgehog ( Hh) signaling pathway inhibitor , on the biological behavior of intrahepatic cholangiocarcinoma cell line RBE .METHODS:The proliferation of RBE cells was detected by cell counting with Typan blue staining and MTT assay , and the apoptosis was analyzed by the flow cytometry . The Transwell invasive cabin assay was used to detect the invasion ability , and Western blot was used to determine the pro-tein expression of Gli 1 and MMP-9 in the RBE cells before and after cyclopamine treatment .RESULTS:Cyclopamine in-hibited the growth of RBE cells in a time-and dose-dependent manner .After cyclopamine treatment for 24 h, 48 h and 72 h, the apoptotic rates were significantly higher than those in control group .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, indica-ting that the invasion ability of the cells declined significantly .Furthermore , Western blot 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:Blockage of the Hh signaling pathway with cyclopamine suppresses the proliferation , promotes the apoptosis and inhibits the invasion ability of RBE cells .