title: Taurolithocholic acid promotes intrahepatic cholangiocarcinoma cell growth via muscarinic acetylcholine receptor and EGFR/ERK1/2 signaling pathway.
abstract: Cholangiocarcinoma (CCA) is a malignant cancer of the biliary tract and its occurrence is associated with chronic cholestasis which causes an elevation of bile acids in the liver and bile duct. The present study aimed to investigate the role and mechanistic effect of bile acids on the CCA cell growth. Intrahepatic CCA cell lines, RMCCA-1 and HuCCA-1, were treated with bile acids and their metabolites to determine the growth promoting effect. Cell viability, cell cycle analysis, EdU incorporation assays were conducted. Intracellular signaling proteins were detected by western immunoblotting. Among eleven forms of bile acids and their metabolites, only taurolithocholic acid (TLCA) concentration dependently (1-40 µM) increased the cell viability of RMCCA-1, but not HuCCA-1 cells. The cell cycle analysis showed induction of cells in the S phase and the EdU incorporation assay revealed induction of DNA synthesis in the TLCA-treated RMCCA-1 cells. Moreover, TLCA increased the phosphorylation of EGFR, ERK 1/2 and also increased the expression of cyclin D1 in RMCCA-1 cells. Furthermore, TLCA-induced RMCCA-1 cell growth could be inhibited by atropine, a non-selective muscarinic acetylcholine receptor (mAChR) antagonist, AG 1478, a specific EGFR inhibitor, or U 0126, a specific MEK 1/2 inhibitor. These results suggest that TLCA induces CCA cell growth via mAChR and EGFR/EKR1/2 signaling pathway. Moreover, the functional presence of cholinergic system plays a certain role in TLCA-induced CCA cell growth.
authors: Chulabhorn Graduate Institute, Bangkok 10210, Thailand.
authors: Sumet
authors: S
authors: Amonyingcharoen
authors: Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok 10210, Thailand.
authors: Tawit
authors: T
authors: Suriyo
authors: Chulabhorn Graduate Institute, Bangkok 10210, Thailand.
authors: Apinya
authors: A
authors: Thiantanawat
authors: Chulabhorn Graduate Institute, Bangkok 10210, Thailand.
authors: Piyajit
authors: P
authors: Watcharasit
authors: Chulabhorn Graduate Institute, Bangkok 10210, Thailand.
authors: Jutamaad
authors: J
authors: Satayavivad
mesh_terms: D001647:Bile Acids and Salts / Q000009:adverse effects* / Q000494:pharmacology; D001650:Bile Duct Neoplasms / Q000378:metabolism / Q000473:pathology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018281:Cholangiocarcinoma / Q000378:metabolism / Q000473:pathology*; D019938:Cyclin D1 / Q000378:metabolism; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D010766:Phosphorylation; D011976:Receptors, Muscarinic; D013658:Taurolithocholic Acid / Q000009:adverse effects* / Q000494:pharmacology
chemical_list: D001647:Bile Acids and Salts; C530104:CCND1 protein, human; D011976:Receptors, Muscarinic; D019938:Cyclin D1; D013658:Taurolithocholic Acid; C512478:EGFR protein, human; D066246:ErbB Receptors