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A study of mifepristone/IFN-γ-induced apoptosis of human cholangiocarcinoma cell line FRH-0201 in vitro

Authors Sun QL, Zhang XG, Xing QT, Ding P, Feng JB, Wu XP, Wang ZM

Published Date November 2012 Volume 2012:5 Pages 335—342

DOI http://dx.doi.org/10.2147/OTT.S36098

Received 19 July 2012, Accepted 7 September 2012, Published 9 November 2012

Qi-Long Sun, Xu-Guang Zhang, Quan-Tai Xing, Peng Ding, Jin-Bo Feng, Xiao-Peng Wu, Zhan-Min Wang

Department of General Surgery, Qilu Hospital of Shandong University, Jinan, China

Objective: To investigate the effects of mifepristone, a progesterone receptor (PR) antagonist, through the proliferation of human cholangiocarcinoma cell line FRH-0201 in vitro and the possible mechanisms involved.
Methods: A two-step addition of poly-HRP anti-mouse immunoglobulin G detection system was used to detect the expression of PR in FRH-0201 cells. After treatments with various concentrations of mifepristone (10, 20, 40, 80, 160, and 320 µmol/L) at various time intervals (24, 48, and 72 hours), the rate of cell inhibition, the rate of cell apoptosis, and the expression of bax/bcl-2/Fas were analyzed with tetrazolium blue (MTT) assay, flow cytometry, reverse transcription polymerase chain reaction and Western blotting. The effect of mifepristone and mifepristone combined with interferon (IFN)-γ-inducing apoptosis on the cells was observed.
Results: Mifepristone remarkably inhibited the proliferation of FRH-0201 cells, which was revealed by MTT assay in a dose- and time-dependent manner. The inhibitory rate gradually increased following the increase of the dosage of mifepristone from a low dosage (10 µmol/L) to a high dosage (320 µmol/L) at different time intervals. Flow cytometry analysis showed mifepristone increased the rate of the FRH-0201 cell-line apoptosis. Notably, the rate of apoptosis increased markedly when the cells were pretreated with IFN-γ and then treated with mifepristone. In addition, mifepristone obviously upregulated bax and Fas expression and downregulated bcl-2 expression.
Conclusion: Mifepristone effectively inhibited the growth of PR-positive human cholangiocarcinoma cell line FRH-0201 in vitro through multiple mechanisms. Mifepristone combined with IFN-γ might therefore induce the apoptosis of the cell line, which is possibly a beneficial clinical scheme for patients suffering from cholangiocarcinoma.

Keywords: cholangiocarcinoma, apoptosis, mifepristone, progesterone, IFN-γ

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