Furthermore, in order to verify the biological phenotype associated with the selected CCA bile acid biomarker, we compared the expression level of farnesoid X receptor (FXR), transmembrane G protein-coupled receptor (TGR5) and sphingosine 1-phosphate receptor 2 (S1PR2) genes by qRT-PCR (Quantitative real-time reverse-transcriptase polymerase chain reaction) analysis after the treatment of CCA cell lines with GCA, TCDCA and control, respectively. This evidence concerns the gene GPBAR1 and cholangiocarcinoma.