title: Triptolide induces apoptotic cell death of human cholangiocarcinoma cells through inhibition of myeloid cell leukemia-1.
abstract: BACKGROUND
  Cholangiocarcinoma (CCA), a devastating neoplasm, is highly resistant to current chemotherapies. CCA cells frequently overexpress the antiapoptotic protein myeloid cell leukemia-1(Mcl-1), which is responsible for its extraordinary ability to evade cell death. Triptolide, a bioactive ingredient extracted from Chinese medicinal plant, has been shown to inhibit cell proliferation and induce apoptosis in several cancers.
  
  
  METHODS
  CCK-8 assay was performed to detect cell survival rate in vitro. DAPI staining and Flow cytometry were used to analyze apoptosis. Western blot was performed to determine the expression levels of caspase-3, caspase-7, caspase-9, PARP, and Mcl-1. Quantitative real-time PCR and immunofluorescence were used to detect the expression levels of Mcl-1. The nude mice xenograft model was used to evaluate the antitumor effect of triptolide in vivo.
  
  
  RESULTS
  Triptolide reduced cell viability in cholangiocarcinoma cell lines in a dose- and time-dependent manner, with IC50 values of 12.6 ± 0.6 nM, 20.5 ± 4.2 nM, and 18.5 ± 0.7 nM at 48 h for HuCCT1, QBC939, and FRH0201 respectively. Triptolide induced apoptosis in CCA cell lines in part through mitochondrial pathway. Using quantitative real-time PCR, western blot and immunofluorescence, we have shown that triptolide downregulates Mcl-1 mRNA and protein levels. Furthermore, triptolide inhibited the CCA growth in vivo.
  
  
  CONCLUSIONS
  Triptolide has profound antitumor effect on CCA, probably by inducing apoptosis through inhibition of Mcl-1. Triptolide would be a promising therapeutic agent for CCA.
authors.forename: Xiwei
authors.initials: X
authors.lastname: Ding
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.forename: Qingshan
authors.initials: Q
authors.lastname: Pei
authors.forename: Jianmei
authors.initials: J
authors.lastname: Pan
authors.forename: Shuling
authors.initials: S
authors.lastname: Huang
authors.forename: Yan
authors.initials: Y
authors.lastname: Yang
authors.forename: Zhu
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Department of Gastroenterology, Drum Tower Hospital, Affiliated to Medical School of Nanjing University, Nanjing, China. 13770755008@126.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lv
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Zou
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018281:Cholangiocarcinoma / Q000188:drug therapy* / Q000235:genetics* / Q000473:pathology; D004224:Diterpenes / Q000008:administration & dosage*; D004852:Epoxy Compounds / Q000008:administration & dosage; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000096:biosynthesis*; D010616:Phenanthrenes / Q000008:administration & dosage*; D023041:Xenograft Model Antitumor Assays
chemical_list: D004224:Diterpenes; D004852:Epoxy Compounds; C579095:MCL1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D010616:Phenanthrenes; C001899:triptolide
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