Cclp1

Four cell lines of cholangiocarcinoma (CCLP1, HuCCT1, RBE, 9810), the bile duct epithelial cell line (HIBEC) cells were obtained from the American Type Culture Collection and were maintained in Dulbecco modified Eagle medium containing 10% (v/v) fetal bovine serum at 37 degree in 5% CO₂ condition. All cell lines were routinely tested to be negative for mycoplasma contamination. Transfection of siRNA and plasmids was performed using Lipofectamine 3000 Reagents (Thermo Fisher Scientific, Waltham, MA) following the manufacturer’s instructions. The sequences of siRNAs against specific targets were listed in Table S1.

The human cholangiocarcinoma cell line FRH–0201, CCLP–1 and the human intrahepatic bile duct epithelial cell line HIBEC, which were all purchased from American Type Culture Collection(ATCC, USA), were cultured in RPMI–1640 (Gibco, USA) supplemented with 10% (v/v) fetal bovine serum (Gibco, USA) at 37°C in a humidified chamber under 5% (v/v) CO2.
Forty matched samples of cancer-adjacent tissue and cholangiocarcinoma tissue were obtained from patients undergoing surgical resection at the Guangdong General Hospital, after written informed consent was obtained from all patients. The matched cancer- adjacent samples were obtained at least 5 cm away from the tumor site. This study was approved by the Research Ethics Committee of Guangdong General Hospital (The approval number was GDREC2015097H).

Human normal bile duct cell line and CCA cell lines HuCCT1, RBE, QBC939, TFK‐1, HCCC9810, Huh‐28, and CCLP1 were purchased from American Type Culture Collection (ATCC) and were cultured in RPMI‐1640 medium (Pricella Life Science & Technology Co., Ltd) supplemented with 10% fetal bovine serum (Scitecher, Beijing Dingguo Changsheng Biotechnology Co., Ltd) at 37°C in a 5% CO₂ incubator.