title: Ursodeoxycholic acid induces apoptosis in hepatocellular carcinoma xenografts in mice.
abstract: OBJECTIVE
  To evaluate the efficacy of ursodeoxycholic acid (UDCA) as a chemotherapeutic agent for the treatment of hepatocellular carcinoma (HCC).
  
  
  METHODS
  BALB/c nude mice were randomized into four groups 24 h before subcutaneous injection of hepatocarcinoma BEL7402 cells suspended in phosphate buffered saline (PBS) into the right flank. The control group (n = 10) was fed a standard diet while treatment groups (n = 10 each) were fed a standard daily diet supplemented with different concentrations of UDCA (30, 50 and 70 mg/kg per day) for 21 d. Tumor growth was measured once each week, and tumor volume (V) was calculated with the following equation: V = (L × W(2)) × 0.52, where L is the length and W is the width of the xenograft. After 21 d, mice were killed under ether anesthesia, and tumors were excised and weighed. Apoptosis was evaluated through detection of DNA fragmentation with gel electrophoresis and the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay. Western blot analysis was performed to determine the expression of apoptosis-related proteins BAX, BCL2, APAF1, cleaved caspase-9, and cleaved caspase-3.
  
  
  RESULTS
  UDCA suppressed tumor growth relative to controls. The mean tumor volumes were the following: control, 1090 ± 89 mm(3); 30 mg/kg per day, 612 ± 46 mm(3); 50 mg/kg per day, 563 ± 38 mm(3); and 70 mg/kg per day, 221 ± 26 mm(3). Decreased tumor volumes reached statistical significance relative to control xenografts (30 mg/kg per day, P < 0.05; 50 mg/kg per day, P < 0.05; 70 mg/kg per day, P < 0.01). Increasing concentrations of UDCA led to increased DNA fragmentation observed on gel electrophoresis and in the TUNEL assay (control, 1.6% ± 0.3%; 30 mg/kg per day, 2.9% ± 0.5%; 50 mg/kg per day, 3.15% ± 0.7%, and 70 mg/kg per day, 4.86% ± 0.9%). Western blot analysis revealed increased expression of BAX, APAF1, cleaved-caspase-9 and cleaved-caspase-3 proteins, which induce apoptosis, but decreased expression of BCL2 protein, which is an inhibitor of apoptosis, following administration of UDCA.
  
  
  CONCLUSIONS
  UDCA suppresses growth of BEL7402 hepatocellular carcinoma cells in vivo, in part through apoptosis induction, and is thus a candidate for therapeutic treatment of HCC.
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Xu
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Yu-Zhen
authors.initials: YZ
authors.lastname: Zhang
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Han
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Tie-Jun
authors.initials: TJ
authors.lastname: Liang
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Wei
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Cheng-Yong
authors.initials: CY
authors.lastname: Qin
authors.affiliation: Hui Liu, Hong-Wei Xu, Ya Huang, Guo-Qing Han, Tie-Jun Liang, Li-Li Wei, Cheng-Yong Qin, Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
authors.forename: Cheng-Kun
authors.initials: CK
authors.lastname: Qin
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D013997:Time Factors; D047368:Tumor Burden / Q000187:drug effects; D014580:Ursodeoxycholic Acid / Q000494:pharmacology*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D014580:Ursodeoxycholic Acid
keywords: Hepatocellular carcinoma; Inhibitory effects; Mechanisms; Ursodeoxycholic acid; Xenografts